CN107337704B - A kind of rare ginsenoside Rk2And Rh3Separation method - Google Patents
A kind of rare ginsenoside Rk2And Rh3Separation method Download PDFInfo
- Publication number
- CN107337704B CN107337704B CN201710450254.7A CN201710450254A CN107337704B CN 107337704 B CN107337704 B CN 107337704B CN 201710450254 A CN201710450254 A CN 201710450254A CN 107337704 B CN107337704 B CN 107337704B
- Authority
- CN
- China
- Prior art keywords
- phase
- ginsenoside
- sample
- mobile phase
- chromatogram
- 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.)
- Expired - Fee Related
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J17/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, having an oxygen-containing hetero ring not condensed with the cyclopenta(a)hydrophenanthrene skeleton
- C07J17/005—Glycosides
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
Ginsenoside Rk is separated with high-speed countercurrent chromatography the present invention relates to a kind of2And ginsenoside Rh3The method of monomer.Solvent condition are as follows: n-hexane-ethyl acetate-methanol-water is as two-phase solvent system, and upper phase is as stationary phase, and lower phase is as mobile phase, and connection type uses just reversed turn, UV detector detection.The isolated rare ginsenoside Rk of the method2And ginsenoside Rh3Purity is all larger than 90%, has many advantages, such as that sample purity is high, sample is lost less, separative efficiency is high.
Description
Technical field
The invention belongs to natural drug separation fields, are related to one kind and isolate and purify ginsenoside Rk2、Rh3Method especially relates to
And a kind of utilization high speed adverse current chromatogram separating and purifying high-purity ginsenoside Rk2、 Rh3Method.
Background technique
Ginsenoside Rk2With good inhibition growth of tumour cell, induction differentiation cancer cell, there is anti-infiltration and anti-rotation
It moves, increase immunity, the performance of the Synergy and attenuation of chemical anticarcinogenic drug, carcinogenesis substance, can be used for radiotherapy, the synergy of chemotherapy subtracts
Poison.Ginsenoside Rh3With antitumaous effect.
Ginsenoside Rk2And Rh3Structural formula it is as follows
Ginsenoside Rk2And Rh3It is general to pass through the hydrolysis common ginsenoside of glycol group such as ginsenoside Rb1、Rb2、Rb3、Rc、
Rd etc. is obtained.But ginsenoside Rk2And Rh3It is isomer, is difficult to separate Rk by common chromatography etc.2、Rh3Monomer,
Rk can be separated using preparative high performance liquid chromatography2、Rh3, but it is low efficiency, time-consuming, and fixed relative sample has irreversible suction
Attached effect, sample loss are big.High speed adverse current chromatogram (HSCCC) is that the one kind to grow up in recent years is continuously not necessarily to stationary phase branch
Efficient, the quick liquid luquid partition chromatography isolation technics of object is supportted, there is big fractional dose, sample nondestructive mistake, rate of recovery height, save
The features such as solvent, has been widely used for preparative separation and the purifying of the fields chemical substances such as biology, medicine, environmental protection.
Summary of the invention
The object of the present invention is to provide prepare Rk using high speed adverse current chromatogram2、Rh3The method of monomer, to overcome the prior art
Present in deficiency.
The invention is realized by the following technical scheme:
It is a kind of to utilize high speed adverse current chromatogram separation preparation Rk2、Rh3The method of monomer is formed by separating and detecting two parts.
Rk used in the present invention2、Rh3Mixture obtains by the following method.Take stem and leaves of American ginseng saponin(e under alkaline condition
It degrades, degradation condition are as follows: sodium hydroxide is put into glycerol and is warming up to 130 DEG C, wherein sodium hydroxide and qualities of glycerin body
Product is than being 1:10；It adds and is heated to 245 DEG C with the stem and leaves of American ginseng saponin(e of the weight such as sodium hydroxide, react 3 hours；Drop
It solves product and separates (mobile phase is ethyl acetate: petroleum ether: ethyl alcohol=100:100:2.3) through silica gel column chromatography, merge and contain someone
Join saponin(e Rk2、Rh3Component, recycling design obtain Rk2、 Rh3Mixture (Rk2、Rh3The sum of content is about 70%).
Rk2、Rh3Isolating and purifying using high speed adverse current chromatogram for mixture carries out, comprising the following steps: with volume ratio 9~
10:13~16:11~12:10 n-hexane-ethyl acetate-methanol-water forms high-speed counter-current solvent system, after being sufficiently mixed
It stands, to upper and lower two phase stratification, takes phase as stationary phase, lower phase is mobile phase, and connection type uses just reversed turn, will fix
It is mutually full of the lattice coil splitter of high-speed counter-current chromatograph, setting high-speed counter-current chromatograph, column temperature is 25~30 DEG C, revolving speed
It for 700~900r/min, 1.5~2.5ml/min of flow rate of mobile phase, is detected in 203nm, after mobile phase flows into, until chromatogram
Baseline keeps stablizing, and takes Rk2、Rh3Sample is mixed, with sample introduction after the dissolution of lower phase solvent, fraction is collected according to chromatogram, is received respectively
Collect ginsenoside Rk2、Rh3Chromatographic peak (Rk2First appearance, Rh3Appearance afterwards is shown in Fig. 1), it is concentrated under reduced pressure, it is dry, Rk is made2、Rh3It is single
Body.
Rk after taking separation respectively2、Rh3Monomer detects its purity, high-efficient liquid phase chromatogram condition with high performance liquid chromatograph
Are as follows: mobile phase acetonitrile-water (50:50), 30 DEG C of chromatographic column temperature, Detection wavelength 203nm.
There is separation method of the invention separation process can be carried out continuously, and Irreversible Adsorption is not present, and have separating effect
The features such as good, solvent usage is few, pollution-free, easy to operate.
Detailed description of the invention
Fig. 1 ginsenoside Rk2、Rh3Chromatographic peak
Specific embodiment
Below with reference to embodiment, the present invention will be further described, but the scope of protection of present invention is not limited to down
Column embodiment.
Embodiment 1
It takes n-hexane 450ml, ethyl acetate 585ml, methanol 495ml, water 450ml to set in 2000ml separatory funnel, vibrates
It shakes up, 15min is stood after layering, oscillation is three times.The upper and lower solution, ultrasonic degassing are filtered respectively with 0.22 μm of filter membrane
30min, it is spare.Mixed sample powder 96mg accurately is weighed, uses 8ml flowing phased soln as sample solution, it is spare.Set instrument
25 DEG C of column temperature, upper phase is injected in tetrafluoroethene pipe with the maximum flow rate of 20ml/min, after being full of upper phase in pipeline, followed by
Multithread goes out phase on 100ml, and connection type is selected just reversed turn, engine speed is adjusted to 850r/min, is led with the flow velocity of 2ml/min
Enter mobile phase, after mobile phase is flowed out from host mouth, then is persistently pumped into 30min and two phase solvent system is allowed to reach dynamic in pipeline
State balances, then baseline held stationary on chromatogram injects 8ml sample solution from injection port, flow velocity is adjusted to 1.5ml/min,
It is detected under 203nm wavelength, each component is collected according to chromatogram, recycling design obtains Rk2、Rh3Monomer.
Take the Rk that above-mentioned steps obtain2、Rh3Monomer sample 5mg is dissolved in 1ml methanol solution, utilizes high-efficient liquid phase color
It composes normalization method and carries out purity check.Mobile phase is acetonitrile-water (50:50), Detection wavelength 203nm.The result shows that Rk2Purity
98.8%, Rh394.4%.
Embodiment 2:
It takes n-hexane 450ml, ethyl acetate 720ml, methanol 540ml, water 450ml to set in 2000ml separatory funnel, vibrates
It shakes up, 15min is stood after layering, oscillation is three times.The upper and lower solution, ultrasonic degassing are filtered respectively with 0.22 μm of filter membrane
30min, it is spare.Mixed sample powder 96mg accurately is weighed, uses 8ml flowing phased soln as sample solution, it is spare.Set instrument
30 DEG C of column temperature, upper phase is injected in tetrafluoroethene pipe with the maximum flow rate of 20ml/min, after being full of upper phase in pipeline, followed by
Multithread goes out phase on 100ml, and connection type is selected just reversed turn, engine speed is adjusted to 850r/min, is led with the flow velocity of 2ml/min
Enter mobile phase, after mobile phase is flowed out from host mouth, then is persistently pumped into 30min and two phase solvent system is allowed to reach dynamic in pipeline
State balances, then baseline held stationary on chromatogram injects 8ml sample solution from injection port, flow velocity is adjusted to 1.5ml/min,
It detects and records under 203nm wavelength, each component is collected according to chromatogram, recycling design obtains Rk2、Rh3Monomer.Take above-mentioned steps
Obtained Rk2、Rh3Monomer sample 5mg is dissolved in 1ml methanol solution, carries out purity using high performance liquid chromatography normalization method
It examines.Mobile phase is acetonitrile-water (50:50), Detection wavelength 203nm.The result shows that Rk2Purity is 95.8%, Rh3Purity is
93.2%.
Claims (3)
1. a kind of rare ginsenoside Rk2And Rh3Separation method, it is characterised in that: take containing Rk2、Rh3Sample, sample point
It is carried out from purification application high speed adverse current chromatogram, with n-hexane-ethyl acetate-methanol-water volume ratio for 9~10:13~16:
11~12:10 forms high-speed counter-current solvent system, takes phase as stationary phase, and lower phase is mobile phase, and high-speed counter-current chromatograph turns
Speed is 700~900r/min, and flow rate of mobile phase 1.5-2.5ml/min, column temperature is 25~30 DEG C, is collected and is flowed according to chromatogram
Part, it is concentrated under reduced pressure, is dry, Rk is made2、Rh3Monomer.
2. separation method as described in claim 1, it is characterised in that high-speed counter-current chromatograph connection type uses just reversed turn.
3. separation method as described in claim 1, it is characterised in that Detection wavelength 203nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710450254.7A CN107337704B (en) | 2017-06-15 | 2017-06-15 | A kind of rare ginsenoside Rk2And Rh3Separation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710450254.7A CN107337704B (en) | 2017-06-15 | 2017-06-15 | A kind of rare ginsenoside Rk2And Rh3Separation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107337704A CN107337704A (en) | 2017-11-10 |
| CN107337704B true CN107337704B (en) | 2019-05-31 |
Family
ID=60221404
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710450254.7A Expired - Fee Related CN107337704B (en) | 2017-06-15 | 2017-06-15 | A kind of rare ginsenoside Rk2And Rh3Separation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107337704B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020215265A1 (en) * | 2019-04-25 | 2020-10-29 | 邦泰生物工程(深圳)有限公司 | Method for preparing mixture of rare ginsenosides rh3 and rk2 and mixture thereof |
| CN111018937A (en) * | 2019-11-25 | 2020-04-17 | 国际竹藤中心 | Method for preparing soapberry saponin monomer by high-speed counter-current chromatography |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101538308A (en) * | 2008-03-21 | 2009-09-23 | 孙成贺 | Method for extracting and preparing high-purity ginsenoside Re from herminium by high speed counter current chromatography |
| CN103193846A (en) * | 2013-04-01 | 2013-07-10 | 中国药科大学 | Preparation method of cis-trans isomers of ginsenoside Rk3 and ginsenoside Rh4 |
| CN105769891A (en) * | 2016-04-12 | 2016-07-20 | 深圳以诺生物制药有限公司 | Low-polarity rare ginsenoside mixture and application thereof |
-
2017
- 2017-06-15 CN CN201710450254.7A patent/CN107337704B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101538308A (en) * | 2008-03-21 | 2009-09-23 | 孙成贺 | Method for extracting and preparing high-purity ginsenoside Re from herminium by high speed counter current chromatography |
| CN103193846A (en) * | 2013-04-01 | 2013-07-10 | 中国药科大学 | Preparation method of cis-trans isomers of ginsenoside Rk3 and ginsenoside Rh4 |
| CN105769891A (en) * | 2016-04-12 | 2016-07-20 | 深圳以诺生物制药有限公司 | Low-polarity rare ginsenoside mixture and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107337704A (en) | 2017-11-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102070690B (en) | Method for preparing adenosine, cordycepin and N6-(2-hydroxyethyl)adenosine simultaneously used as chemical reference substances | |
| CN103724389B (en) | The method of antineoplastic component ganoderic acid C 1 and Ganodenic acid F is prepared in a kind of separation | |
| CN103145677B (en) | Method for separating active ingredients from aquilaria sinensis lamina by utilizing high-speed countercurrent chromatography | |
| CN104003963B (en) | A kind of method for separating and preparing of cnidium lactone | |
| CN108129467A (en) | A kind of HSCCC-DPPH is combined online activity analysis and the method for detaching Radix Ophiopogonis homoisoflavone | |
| CN107337704B (en) | A kind of rare ginsenoside Rk2And Rh3Separation method | |
| CN106349301B (en) | Method for separating and purifying punicalagin in pomegranate peel | |
| CN105367531A (en) | Method for separating two homoisoflavonoids in fibrous roots of ophiopogon japonicusby adopting recycling high-speed counter-current chromatography | |
| CN104829666A (en) | Method for preparing high purity baicalin from radix scutellariae | |
| CN114988979B (en) | Method for preparing high-purity lycopene by macro separation | |
| CN102617674B (en) | Preparation method of scopolin monomer in anisodus tanguticus root | |
| CN109053636B (en) | Method for preparing epoxy carrot olefine aldehyde A and B | |
| CN100500689C (en) | Method of separating and preparing wild jujube seed saponin | |
| CN108329292A (en) | A method of preparing former haematoxylin B | |
| CN109096078A (en) | A kind of preparation method of macrocarpal C | |
| CN108675915A (en) | A method of preparing ent-spathulenol | |
| CN108546261A (en) | A method of preparing protosappanin A | |
| CN107556275B (en) | Preparation method of atractylenolide II | |
| CN102127124B (en) | Method for preparing hydroxysafflor yellow A | |
| CN102911240A (en) | Method for preparing high-purity bayogenin | |
| CN108546260A (en) | A method of preparing former haematoxylin C | |
| CN108675916A (en) | It is a kind of to prepare aromadendrane-4 β, the method for 10 α-diol | |
| CN108623434A (en) | A method of preparing Alismoxide | |
| CN103896892A (en) | Preparation of senkyunolide I and senkyunolide H from ligusticum wallichii extract via high-speed countercurrent chromatography | |
| CN102250169A (en) | Preparation method for frangulin B |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Liu Songyan Inventor after: Zhao Pengyun Inventor after: Jin Yongri Inventor after: Li Xuwen Inventor before: Jin Yongri Inventor before: Zhao Pengyun Inventor before: Li Xuwen Inventor before: Liu Songyan |
|
| CB03 | Change of inventor or designer information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190531 Termination date: 20200615 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |