CN105294629A - Casticin preparation method - Google Patents
Casticin preparation method Download PDFInfo
- Publication number
- CN105294629A CN105294629A CN201510795076.2A CN201510795076A CN105294629A CN 105294629 A CN105294629 A CN 105294629A CN 201510795076 A CN201510795076 A CN 201510795076A CN 105294629 A CN105294629 A CN 105294629A
- Authority
- CN
- China
- Prior art keywords
- vitexicarpin
- preparation
- extract
- described step
- casticin
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/40—Separation, e.g. from natural material; Purification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to a casticin preparation method, which comprises: (1) adding crushed fructus viticis raw material to ethanol, carrying out ultrasonic extraction, concentrating the extraction solution to obtain an extract, dissolving the extract with an alcohol aqueous solution according to a solid-liquid mass ratio, extracting, and carrying out pressure reducing concentration to obtain an extract; (2) eluting the extract with an alcohol aqueous solution by using column chromatography and by adopting polyamide as a filler, collecting the fraction containing casticin, concentrating, and drying to obtain a crude extract; and (3) separating the crude extract by using a high-speed countercurrent chromatography method, carrying out online monitoring through an ultraviolet detector, collecting the fraction, carrying out pressure reducing concentration, crystallizing, and drying to obtain the casticin, wherein the solvent system of the high-speed countercurrent chromatography comprises n-hexane, ethyl acetate, methanol and water. The casticin preparation method of the present invention has characteristics of good product purity and good product quality, is suitable for various types of high-speed countercurrent chromatographs, and is suitable for industrialization amplification.
Description
Technical field
The invention belongs to natural drug extraction and isolation field, particularly a kind of preparation method of Vitexicarpin.
Background technology
Simpleleaf Shrub Chastetree Fruit is the dry mature fruit of Verbenaceae Vitex Vitex rotundifolia VitextrifoliaL.var.simplicifoliaCharm. or wild pepper VitextrifoliaL., mainly originates in the ground such as Shandong, Guangxi, Yunnan, Jiangxi, Zhejiang, Fujian and Guangdong.Simpleleaf Shrub Chastetree Fruit is conventional Chinese medicine, has effect of dispelling wind and heat pathogens, the clear sharp head, pain relieving.Be used for the treatment of the diseases such as wind-heat, cold headache, migraine clinically.It is reported, the main active ingredient of Simpleleaf Shrub Chastetree Fruit is flavonoid compound, and wherein Vitexicarpin is as the quality control index of the Viticis Fructus of China's 2010 editions States Pharmacopoeia specifications.
Present research shows, Vitexicarpin has anti-hypoxia, anti-pessimal stimulation, antifatigue, two-ways regulation body, suppresses blood sugar increasing and affect the multiple effects such as learning and memory.As can be seen here, the exploitation of target component has commercial value widely.Structural formula is as follows:
At present, the Vitexicarpin extraction and isolation in Simpleleaf Shrub Chastetree Fruit mainly more adopts column chromatography, and separation cycle is long, consumes solvent many, complicated operation.High speed adverse current chromatogram (High-speedcountercurrentchromatography, HSCCC) as a kind of novel separating and purifying technology, easy to operate, fast, resolution is higher, overcome the shortcoming such as sample combination, inactivation, pollution brought by solid phase carrier, be widely used in the separation preparation of natural product active ingredient and the Analysis and Identification of total herbal medicine.Have bibliographical information high speed adverse current chromatogram separation and purification Vitexicarpin, but have applied sample amount few, the shortcomings such as disengaging time is long, and purification effect is bad, can not accomplish scale production.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of preparation method of Vitexicarpin, and the method prepares Vitexicarpin, and product purity is high, quality better, is applicable to various model high-speed counter-current chromatograph, is easy to industrialization and amplifies.
The preparation method of a kind of Vitexicarpin of the present invention, comprising:
(1) by the Simpleleaf Shrub Chastetree Fruit raw material after pulverizing, add ethanolic soln, supersound extraction with solid-liquid mass ratio 1:5-10, concentrated extracting solution obtains medicinal extract (being evaporated to without alcohol taste); Dissolved by solid-liquid mass ratio 1:100-150 alcohol solution by medicinal extract, extraction, concentrating under reduced pressure is extracted thing;
(2) adopting column chromatography, take polymeric amide as filler, the above-mentioned extract of alcohol solution wash-out, collects the stream part containing Vitexicarpin, concentrated, dry, obtains crude extract;
(3) adopt high-speed countercurrent chromatography to be separated to above-mentioned crude extract, the solvent systems of high speed adverse current chromatogram by normal hexane, ethyl acetate, first alcohol and water by volume 1:1-2:1-2:2 form; UV-detector on-line monitoring, collects stream part concentrating under reduced pressure, crystallization, dry, obtains Vitexicarpin.
The volume fraction of the ethanol in described step (1) is 30-90%.
Supersound extraction number of times in described step (1) is 2 ~ 3 times.
Alcohol solution in described step (1) is the aqueous ethanolic solution of volume fraction 30-90%.
Extraction solvent for use in described step (1) is sherwood oil.
Alcohol solution in described step (2) is the aqueous ethanolic solution of volume fraction 75-95%.
The separation condition of the high speed adverse current chromatogram in described step (3) is: engine speed 750-1000rpm/min, moving phase 2.5-15.0ml/min, bath temperature 10-35 DEG C.
The upper of solvent systems in described step (3) is stationary phase mutually, and lower is moving phase mutually.
Sample is carried out special easy pre-treatment by the present invention, and make sample purity improve 1 times, applied sample amount increases by 3 times, gropes solvent system simultaneously, makes disengaging time shorten 1 times, is intended to solve existing separating technology efficiency lower, fails to realize the problem of a large amount of preparative separation.
beneficial effect
(1) separation method of the present invention is easy, and fast, separation cycle is short, can be separated preparation high purity Vitexicarpin in the short period of time.Traditional column chromatography methods complex operation, need repeatedly carry out column chromatography for separation, separation cycle is long, and target component extremely adsorbs seriously.
(2) monomer preparation amount of the present invention is large, according to the quality of required Vitexicarpin, can regulate the sample size of separated sample, and adopt countercurrent chromatography instrument, technique favorable reproducibility, product purity is high, has a good application prospect.
Accompanying drawing explanation
Fig. 1 is that embodiment 1HSCCC is separated the Vitexicarpin collection of illustrative plates obtained;
Fig. 2 is Vitexicarpin HPLC collection of illustrative plates.
Embodiment
Below in conjunction with specific embodiment, set forth the present invention further.Should be understood that these embodiments are only not used in for illustration of the present invention to limit the scope of the invention.In addition should be understood that those skilled in the art can make various changes or modifications the present invention, and these equivalent form of values fall within the application's appended claims limited range equally after the content of having read the present invention's instruction.
Embodiment 1
Learn from else's experience dry Simpleleaf Shrub Chastetree Fruit raw material pulverizing, claim 2000g, add 10L80% ethanolic soln supersound extraction 60 minutes, extract 3 times, concentrating under reduced pressure extracting solution obtains medicinal extract 45g at every turn.Add 100 mass parts 50% aqueous ethanolic solutions by 1 mass parts medicinal extract to dissolve, petroleum ether extraction, coextraction 3 times, combining extraction liquid reclaims sherwood oil and obtains extract 40g, on extract, order number is the polyamide column of 100-200, with the aqueous ethanolic solution wash-out of volume fraction 80%, collects elutriant, concentrate drying obtains crude extract, is separated for high-speed counter-current.
Normal hexane, ethyl acetate, methyl alcohol, water two phase solvent system is prepared, 1:2:1:2 mixing by volume, stratification after jolting, ultrasonic degas 30min in separating funnel.Get and fill high-speed counter-current chromatograph mutually as stationary phase, rotate main frame, rotating speed 800rpm, pump into 5ml/min simultaneously and do moving phase mutually down, bath temperature is 20 DEG C, after setting up running balance, crude extract is dissolved by moving phase, by sampling valve sample introduction, UV-detector is on-line monitoring under 254nm wavelength, collects each target component, continuous production, reagent is reclaimed in corresponding merging, methanol crystallization after concentrated, is separated and obtains highly purified Vitexicarpin 200mg from 1000mg crude extract.Detect through HPLC, content 98.0%, the data characterizing its physical behavior through UV, IR, MS, HNMR, CNMR etc. are consistent with prior art.
Embodiment 2
Learn from else's experience dry Simpleleaf Shrub Chastetree Fruit raw material pulverizing, claim 2000g, add 10L95% ethanolic soln supersound extraction 60 minutes, extract 3 times, concentrating under reduced pressure extracting solution obtains medicinal extract 40g at every turn.Add 100 mass parts 50% aqueous ethanolic solutions by 1 mass parts medicinal extract to dissolve, petroleum ether extraction, coextraction 3 times, combining extraction liquid reclaims sherwood oil and obtains extract 35g, on extract, order number is the polyamide column of 200-300, with the aqueous ethanolic solution wash-out of volume fraction 95%, collects elutriant, concentrate drying obtains crude extract, is separated for high-speed counter-current.
Normal hexane, ethyl acetate, methyl alcohol, water two phase solvent system is prepared, 1:2:2:2 mixing by volume, stratification after jolting, ultrasonic degas 30min in separating funnel.Get and fill high-speed counter-current chromatograph mutually as stationary phase, rotate main frame, rotating speed 800rpm, pump into 10ml/min simultaneously and do moving phase mutually down, bath temperature is 25 DEG C, after setting up running balance, crude extract is dissolved by moving phase, by sampling valve sample introduction, UV-detector is on-line monitoring under 254nm wavelength, collects each target component, continuous production, reagent is reclaimed in corresponding merging, methanol crystallization after concentrated, is separated and obtains highly purified Vitexicarpin 300mg from 1000mg crude extract.Detect through HPLC, content 97.7%, the data characterizing its physical behavior through UV, IR, MS, HNMR, CNMR etc. are consistent with prior art.
Embodiment 3
Learn from else's experience dry Simpleleaf Shrub Chastetree Fruit raw material pulverizing, claim 2000g, add 10L70% ethanolic soln supersound extraction 60 minutes, extract 3 times, concentrating under reduced pressure extracting solution obtains medicinal extract 60g at every turn.Add 100 mass parts 70% aqueous ethanolic solutions dissolvings by 1 mass parts medicinal extract and add petroleum ether extraction, coextraction 3 times, combining extraction liquid reclaims sherwood oil and obtains extract 51g, on extract, order number is the polyamide column of 300-400, with the aqueous ethanolic solution wash-out of volume fraction 75%, collect elutriant, concentrate drying obtains crude extract, is separated for high-speed counter-current.
Normal hexane, ethyl acetate, methyl alcohol, water two phase solvent system is prepared, 1:2:1:1 mixing by volume, stratification after jolting, ultrasonic degas 30min in separating funnel.Get and fill high-speed counter-current chromatograph mutually as stationary phase, rotate main frame, rotating speed 800rpm, pump into 8ml/min simultaneously and do moving phase mutually down, bath temperature is 30 DEG C, after setting up running balance, crude extract is dissolved by moving phase, by sampling valve sample introduction, UV-detector is on-line monitoring under 254nm wavelength, collects each target component, continuous production, reagent is reclaimed in corresponding merging, methanol crystallization after concentrated, is separated and obtains highly purified Vitexicarpin 250mg from 1000mg crude extract.Detect through HPLC, content 98.8%, the data characterizing its physical behavior through UV, IR, MS, HNMR, CNMR etc. are consistent with prior art.
Claims (8)
1. a preparation method for Vitexicarpin, comprising:
(1) by the Simpleleaf Shrub Chastetree Fruit raw material after pulverizing, add ethanolic soln, supersound extraction with solid-liquid mass ratio 1:5-10, concentrated extracting solution obtains medicinal extract; Dissolved by solid-liquid mass ratio 1:100-150 alcohol solution by medicinal extract, extraction, concentrating under reduced pressure is extracted thing;
(2) adopting column chromatography, take polymeric amide as filler, the above-mentioned extract of alcohol solution wash-out, collects the stream part containing Vitexicarpin, concentrated, dry, obtains crude extract;
(3) adopt high-speed countercurrent chromatography to be separated to above-mentioned crude extract, the solvent systems of high speed adverse current chromatogram by normal hexane, ethyl acetate, first alcohol and water by volume 1:1-2:1-2:2 form; UV-detector on-line monitoring, collects stream part concentrating under reduced pressure, crystallization, dry, obtains Vitexicarpin.
2. the preparation method of a kind of Vitexicarpin according to claim 1, is characterized in that: the volume fraction of the ethanol in described step (1) is 30-90%.
3. the preparation method of a kind of Vitexicarpin according to claim 1, is characterized in that: the supersound extraction number of times in described step (1) is 2 ~ 3 times.
4. the preparation method of a kind of Vitexicarpin according to claim 1, is characterized in that: the alcohol solution in described step (1) is the aqueous ethanolic solution of volume fraction 30-90%.
5. the preparation method of a kind of Vitexicarpin according to claim 1, is characterized in that: the extraction solvent for use in described step (1) is sherwood oil.
6. the preparation method of a kind of Vitexicarpin according to claim 1, is characterized in that: the alcohol solution in described step (2) is the aqueous ethanolic solution of volume fraction 75-95%.
7. the preparation method of a kind of Vitexicarpin according to claim 1, it is characterized in that: the separation condition of the high speed adverse current chromatogram in described step (3) is: engine speed 750-1000rpm/min, moving phase 2.5-15.0ml/min, bath temperature 10-35 DEG C.
8. the preparation method of a kind of Vitexicarpin according to claim 1, is characterized in that: the upper of the solvent systems in described step (3) is stationary phase mutually, and lower is moving phase mutually.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510795076.2A CN105294629A (en) | 2015-11-18 | 2015-11-18 | Casticin preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510795076.2A CN105294629A (en) | 2015-11-18 | 2015-11-18 | Casticin preparation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105294629A true CN105294629A (en) | 2016-02-03 |
Family
ID=55192545
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510795076.2A Pending CN105294629A (en) | 2015-11-18 | 2015-11-18 | Casticin preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105294629A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105801593A (en) * | 2016-03-31 | 2016-07-27 | 上海同田生物技术股份有限公司 | Decursitin preparing method |
-
2015
- 2015-11-18 CN CN201510795076.2A patent/CN105294629A/en active Pending
Non-Patent Citations (6)
| Title |
|---|
| 刘亚英: "《蔓荆子的有效成分和质量研究》", 《中国优秀博硕士学位论文全文数据库（硕士） 医药卫生科技辑》 * |
| 吴闯等: "《蔓荆子化学成分研究》", 《药物生物技术》 * |
| 管仁军: "《蔓荆子化学成分及其含量测定方法研究》", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 管仁军等: "《高速逆流色谱分离纯化蔓荆子中的活性成分》", 《色谱》 * |
| 罗娅君等: "《高速逆流色谱分离与鉴定牡荆化学成分》", 《分析测试学报》 * |
| 赵利新: "《单叶蔓荆叶化学成分及蔓荆子的质量控制研究》", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105801593A (en) * | 2016-03-31 | 2016-07-27 | 上海同田生物技术股份有限公司 | Decursitin preparing method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103665060B (en) | The separation purification method of a kind of crocin I monomer, crocin II monomer | |
| CN105294628B (en) | A kind of method that separation from chrysanthemum indicum prepares flavones ingredient | |
| CN103833803A (en) | Method for ultrasonic extracting and resin purifying sweetening agent in Lithocarpus polysachyus rehd leaf | |
| CN103709152B (en) | A kind of separation purification method of puerarin | |
| CN102898347A (en) | Method for extracting caragana microphylla from hypaphorine | |
| CN105367424A (en) | Method for preparing high-purity chlorogenic acid from eupatorium adenophorum Spreng | |
| CN105294629A (en) | Casticin preparation method | |
| CN103694096B (en) | Separation and purification method for ingenol monomers | |
| CN102895303A (en) | Resin purification method for extracting and separating total flavone of cyclocarya paliurus | |
| CN105330710A (en) | Method for simultaneously extracting three kinds of iridoid glycoside chemical reference substances from fructus ligustri lucidi | |
| CN105175426A (en) | Method of extracting and purifying bergenin from cissus pteroclada hayata | |
| CN101618052A (en) | Process for extracting total flavonoids from hippophae leaves | |
| CN105273015B (en) | A kind of preparation method of high-purity Paeoniflorin and albiflorin | |
| CN103275054A (en) | Method for preparing bellidifolin | |
| CN102274254B (en) | Preparation method of alligator alternanthera effective fraction extract | |
| CN102399252A (en) | Preparation method of cowherb seed flavonoid glycoside monomer | |
| CN103232469A (en) | Method for preparing mother chrysanthemum lactone | |
| CN103254164A (en) | Preparation method of atractylenolide I | |
| CN107746419B (en) | A kind of apple flower active component, preparation method and the method for extracting Kaempferol glucoside using it | |
| CN103242399A (en) | Method for extracting apigenin-7-O-beta-D-glucuronide from broussonetia papyrifera leaves | |
| CN103254045A (en) | Preparation method of high-purity Erianin | |
| CN102993143A (en) | Method for rapidly separating alkannic acid monomer from salviae miltiorrhizae | |
| CN103232470A (en) | Method for preparing high-purity ailanthone | |
| CN103421017A (en) | Preparation method of high-purity morusin | |
| CN104447642A (en) | Chromatographic separation and purification method for industrially preparing andrographolide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160203 |
|
| RJ01 | Rejection of invention patent application after publication |