CN100434135C - Method for separating and purifying natural product using three-phase counter current chromatograph - Google Patents
Method for separating and purifying natural product using three-phase counter current chromatograph Download PDFInfo
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Abstract
The present invention relates to a method for separating and purifying natural product by using three-phase counter-current chromatography. Sad method is characterized by that in the immobile phase of conventional counter-current chromatography a solid separation medium-microsphere can be added so as to form a three-phase separation system including liquid immobile phase, solid immobile phase and liquid mobile phase, so that the counter-current chromatography and column chromatography can be combined into one body, therefore the separation and purification efficiency of natural product can be greatly raised.
Description
Technical field
The present invention relates to a kind of method of using three-phase counter-current chromatographic separation and purification natural products.
Background technology
In recent years, to natural products (comprising Chinese medicine)---the research of active component has become the focus of pharmacy, pharmacology and clinical practice in natural plants, animal and the mineral, the prescription medicine in the whole world 25% is from natural plants, wherein existing 121 kinds of reactive compounds can be used as the conventional medicine that uses, and have caused the attention of countries in the world.
China is vast in territory, is one of the abundantest country of plant resources in the world, kind surplus only higher plant just has 30,000, and wherein kind surplus the medicinal plant 11,000 has clinical practice history in several thousand.But the Chinese medicine of China coverage rate in the international market is very little, can not show a candle to neighbouring country Japan.In 210 granted Chinese prescription medicine preparations that Japan has, its prescription mainly is treatise on Febrile Diseases and " Synopsis Golden Chamber " from China well-known doctor Zhang Zhongjing works, and 75% of said preparation raw material also is by China's input, but its covering in the international market has reached 80%.By contrast, China has 4000 kinds of Chinese medicine preparations, but coverage rate in the international market only is 3～5%.Though causing the one of the main reasons of this situation is that China has certain progress to the technology that natural products carries out medicinal production and processing, also there is a big difference from requirement that modern medicine industry high quality is produced.
The natural products composition is very complicated, and active constituent content is very little, removes invalid or even harmful components wherein, reduces toxic and side effect; Improve active constituent content, strengthen curative effect of medication; Forming production technology standard, high quality is to need one of key issue that solves in China's natural drug development process.
The production technology of natural products mainly comprises to be extracted and purifying two parts.
Usually use traditional extracting methods such as solvent extraction method, steam distillation, sublimed method, ultrasonic extraction, or the extracting method that just grows up in recent years such as Microwave Extraction method (MAE) and supercritical carbon dioxide extraction method (SFE), active component (or active principle) enrichment quickly and efficiently with plant is prepared into CE.And then this natural products carried out further separation and purification.
It is technical bottleneck problem in the natural products research that extract is carried out purifying.Purification process commonly used at present comprises classical solvent extraction, fractionating process, the precipitation method, sublimed method, crystallisation, membrane separation process, molecularly distilled, SPE, adverse current chromatogram and column chromatography etc., every kind of method has the advantage of oneself but also has defective and limitation.Wherein, more commonly used with column chromatography and adverse current chromatogram method especially.
Adverse current chromatogram (High-speed counter-current chromatography, HSCCC) be a kind of liquid luquid partition chromatography technology that does not have solid-state supporter, with light and handy polytetrafluoroethylhelix helix tube as splitter, fixing all is liquid with flowing mutually mutually, in the time of the helix tube rotation, do revolution motion around an outside axis that is parallel to the axis of rotation, so as to obtaining enough strong centrifugal force field, under the prerequisite that is kept mutually in two-phase, form to fix and cut apart trend and convection current trend with mobile two-phase mutually, carry out this cutting apart with the process of convection current continuously, if the inlet of the sample that will separate from spiral tube injected, continuous distribution and transmission process will be carried out in tubing string, thereby realizes continuous, liquid-liquid assigning process efficiently.The advantage of this method is: adverse current chromatogram does not have solid phase carrier, has avoided sample to be separated and surface of solid phase carriers to produce chemical reaction and sex change and Irreversible Adsorption; Sample does not need strict preliminary treatment, and at analysiss/separating high-viscosity with easily cause to fix and have clear superiority aspect the sample that adsorbs mutually; The scope of application is more extensive, and organic molecules such as both separable alkaloid, flavones also can isolated polypeptide, macromolecular compound such as protein, polysaccharide; As the natural drug method of quality control, can clearly express in this complex system of Chinese medicine the overall state of various chemical composition CONCENTRATION DISTRIBUTION between different batches, stability and favorable reproducibility; Because of multiple model is arranged, both can realize the analysis of sample, also can carry out scale preparation; Instrumentation is very simple, and it is very easy to change the phase that flows; Its instrument and consumptive material domesticize fully, and cost is starkly lower than high performance liquid chromatography.But this method is owing to the limitation of extract and separate itself, and generally separative efficiency is lower than high performance liquid chromatography.
Column chromatography method has been obtained huge progress after the last century the nineties, possessed higher separating power, selectivity, versatility, and operating condition is relatively gentleer.The column chromatography technology kind is more, and is for example following several:
Adsorption chromatography: this is a kind of absorption of broad sense, not only comprises the physical action of the no chemical bond introducing of common indication, also comprises hydrophobic interaction and hydrogen bond force etc.According to each component in the extract to the fixing difference of phase degree of absorption, with and corresponding flow mutually in the difference of solubility realize.Related medium scope is very extensive, comprising: silica gel, bonding phase silica gel, agarose, glucan, aluminium oxide, macroporous absorbent resin, polyamide, activated carbon, hydroxyapatite etc.Separating ranges is extensive, has both comprised fat-soluble compound, also comprises water soluble compound.
Ion-exchange chromatography: to be charged object molecule combine with the ion-exchange group of oppositely charged on the salt of other band identical charges or charged impurity molecule competition ground and the medium its isolation technics, thus since sample molecule with exchange the different realizations of the interactional intensity in site and separate.Can the separating water-soluble composition, as amino acid, peptide class, alkaloids and polyphenol compound etc.
Perfusion chromatography: used the special chromatography media that has through hole, greatly reduced resistance to mass tranfer, shortened the mass transfer approach, improved post and imitated and separating rate.
The application of column chromatography for separation efficient height, especially high performance liquid chroma-tography, its stationary-phase particle size diameter is little, and total surface area is big, therefore has very high resolution capability and good selectivity; The compounds suitable for use type is extensive.Thereby but also exist post absorption to cause sample loss, and the mesolow column chromatography for separation time is long, and part medium costliness makes separation costs height, the compound of some structural similarity compound that especially hydrophily is strong adopt one-step method can't realize problems such as effectively separation.
Summary of the invention
Overall resolution is lower when the objective of the invention is to overcome use adverse current chromatogram method separating natural product, and optionally limit because of it when using column chromatography method separating natural product, adopt one-step method can't realize the compound of some structural similarity, especially the defective of effective separation of the compound that hydrophily is strong, thus a kind of solid-state separating medium with adverse current chromatogram method and column chromatography method is provided---that microballoon combines, as to use three-phase counter-current chromatographic separation and purification natural products method.
The objective of the invention is to realize by the following technical solutions:
The method of use three-phase counter-current chromatographic separation and purification natural products provided by the invention, be the adverse current chromatogram of routine fixing mutually in, add solid-state separating medium---microballoon, form one and comprise the fixedly piece-rate system of phase, solid-state fixedly phase and the liquid three-phase mutually that flows of liquid state, method with adverse current chromatogram method and column chromatography method combine specifically comprises following step:
1) select conventional adverse current chromatogram dicyandiamide solution to flow mutually as the liquid state of three-phase counter-current chromatogram is mutually fixing and liquid, and in separatory funnel balance 2h, two-phase is separated up and down, and is standby;
Described dicyandiamide solution is to go up based on organic facies, polarity a little less than; Following to water, polarity is stronger; Comprise: n-hexane-alcohol-water, n-hexane-ethyl acetate-methanol-water, n-hexane-ethyl acetate-methyl alcohol-acetate-water, ethyl acetate-n-butanol-acetonitrile-water, ethyl acetate, alcohol and water, ethyl acetate-ethanol-acetate-water, ethyl acetate-n-butanol-water, n-butanol-water or n-butanol-acetate-water;
2) the fixedly phase of preparation three-phase counter-current chromatogram is with mutually mobile:
Above-mentioned adverse current chromatogram dicyandiamide solution Semi-polarity is fixed a mutually close phase as liquid fixedly phase with solid-state, be beneficial to stable and even distribute of media particle in this liquid state is fixed mutually; Then with solid-state fixedly be added to above-mentioned liquid state fixing mutually in, mix, the suspension that makes is as the fixedly phase of three-phase counter-current chromatogram;
Described solid-state fixing is lipophilic medium or hydrophilic medium mutually; The solid-state of adding is 2～6g/L with the fixing ratio mutually of liquid state fixedly;
Described lipophilic medium is the cross-linked copolymer of inverse bonded phase silica gel, polystyrene, polyamide or styrene-divinylbenzene; This solid-state fixedly phase lipophilicity is stronger, adds last phase (organic facies) in the dicyandiamide solution of above-mentioned adverse current chromatogram as the fixedly phase of three-phase counter-current chromatogram;
Described hydrophilic medium is positive bonding phase silica gel, agarose matrix gel or glucan matrix gel; This solid-state fixedly phase hydrophily is stronger, adds following phase (water) in the dicyandiamide solution of above-mentioned adverse current chromatogram as the fixedly phase of three-phase counter-current chromatogram;
A dicyandiamide solution Semi-polarity and a solid-state fixing close mobile phase as the three-phase counter-current chromatogram with above-mentioned adverse current chromatogram;
3) set up three alternate balances:
With step 2) obtain contain lipophilic medium fixedly with the splitter (polytetrafluoroethylene (PTFE) twines the helix tube that forms) of the full conventional adverse current chromatogram of the flow pump of 8～10mL/min, then with the rotating speed forward running adverse current chromatogram main frame of 300～600r/min; Pump into mobile phase with low flow velocity 0.5～1.5mL/min simultaneously; Add the back-pressure of 0.7Mpa at the port of export of adverse current chromatogram, wait to flow when overfall overflows, three alternate balances are set up; Under Archimedes spiral power, planetary centrifugal force field and mobile comprehensive function mutually, 25～70% fix is retained in the splitter mutually;
Or with step 2) obtain contain hydrophilic media fixedly with the splitter (polytetrafluoroethylene (PTFE) twines the helix tube that forms) of the full conventional adverse current chromatogram of the flow pump of 8～10mL/min, then with the rotating speed antiport adverse current chromatogram main frame of 300～600r/min; Pump into mobile phase with low flow velocity 0.5～1.0mL/min simultaneously; Add the back-pressure of 1Mpa at the port of export of adverse current chromatogram, wait to flow when overfall overflows, three alternate balances are set up; Under Archimedes spiral power, planetary centrifugal force field and mobile comprehensive function mutually, 25～60% fix is retained in the splitter mutually;
4) with the natural products sample introduction of purifying to be separated:
With the natural products of purifying to be separated by the concentration of 1～10mg/mL be dissolved in flow mutually after, push the sample introduction circle;
5) separation and purification:
In the natural products sample of purifying to be separated each component in liquid state fixing and flow mutually between in the distribution, also adsorption chromatography take place mutually separate with solid-state fixing; Use the online detection cut of UV-detector, and by recorder or computer software record chromatogram, and, obtain the natural products of purifying according to chromatogram collection cut.
In the method for use three-phase counter-current chromatographic separation and purification natural products provided by the invention, constitute the liquid two-phase and a solid-state phase of three-phase,, be suitable for separating different types of compound because of its different in kind.Contain the fixing three-phase that constitutes mutually of lipophilic medium, be suitable for separating the compound of low pole and middle polarity; Contain the fixing three-phase that constitutes mutually of hydrophilic media, be suitable for separating the compound of middle polarity and strong polarity.
The method of use three-phase counter-current chromatographic separation and purification natural products provided by the invention combines adverse current chromatogram method and column chromatography method first.Comprise solid-state fixedly phase, liquid fixing phase and the liquid three-phase system that flows mutually in this method, under certain rotating speed, under Archimedes spiral power, planetary centrifugal force field and mobile comprehensive function mutually, form fixing phase (comprising that liquid fixing phase is with solid-state mutually fixing) with mobile mutually cut apart trend and convection current trend.Adsorbing separation also takes place with solid-state fixing when distributing in each component mutually in the sample between liquid state is fixed and flowed mutually, forms distribution and goes on foot piece-rate system with absorption symphyogenetic one.The transmission and the distribution of the media particle vibration in the planetary centrifugal helix tube of rotation enhancing material, formed theoretical cam curve both had been higher than adverse current chromatogram, also was higher than the low pressure adsorption chromatography; Systemic resolution improves; The retention time of cut prolongs, and helps improving cut purity; Two steps separated and to be reduced to a step and to finish.
When the separation and purification that is applied to natural products, by adjusting the kind of dicyandiamide solution and media particle, realize the combination of multiple choices, in a piece-rate system, realize extract and separate and adsorbing separation under the high speed centrifugation force field, in a running, thereby with the selectivity of two kinds of chromatographic processes in conjunction with the change piece-rate system, one step of separation process of two kinds of principles is finished, solved natural products because of the component complexity, structure proximate and the separation difficulty that causes, the problem that separating degree is low, for of a great variety, the component complexity, structure proximate, the separation of the natural products that active constituent content is low provides new technology, and has solved the lower problem of adverse current chromatogram overall resolution.Environmental protection and national sustainable development strategy have also been taken into account simultaneously.
Description of drawings
Fig. 1 is that embodiment 1 uses the three-phase counter-current chromatographic system to separate root of large-flowered skullcap water extract, uses the collection of illustrative plates that carries out online detection under the UV-detector 285nm.
Fig. 2 is that embodiment 2 uses the three-phase counter-current chromatographic system to separate red sage root fat-soluble compound, uses the collection of illustrative plates that carries out online detection under the UV-detector 280nm.
The specific embodiment
The separation and purification of embodiment 1, root of large-flowered skullcap water extract
Use agarose matrix gel Superose 12 and n-hexane-ethyl acetate-methyl alcohol-acetate-water (volume ratio is 1-6-1.5-1.5-6) system to constitute the three-phase of three-phase counter-current chromatographic system of the present invention, come separation and purification root of large-flowered skullcap water extract, concrete steps are:
1) according to 1: 6: 1.5: 1.5: 6 volume ratio, n-hexane, ethyl acetate, methyl alcohol, acetate and water are mixed, and in separatory funnel balance 2h, mixed liquor is divided into two-phase up and down, obtains n-hexane-ethyl acetate-methyl alcohol-acetate-aqueous solvent system; To be separated up and down;
2) use Superose 12 gels (GE Healthcare, average diameter of particles 10 μ m) as solid-state fixedly phase; With the dicyandiamide solution Semi-polarity in the step 1) with solid-state fixing mutually close following as the fixing phase of liquid state; With Superose 12 gels and solution mixing mutually down, the addition of Superose 12 gels is 2g/L with the following ratio of solution mutually; Speed with 60r/min mixes liquid, and Superose 12 gels are uniformly distributed in down in the phase solution, obtains the fixedly phase of three-phase counter-current chromatographic system of the present invention, comprising solid-state fixedly phase with liquid fixing mutually;
With dicyandiamide solution Semi-polarity in the step 1) and solid-state fixing close last mobile phase as the three-phase counter-current chromatogram;
3) with step 2) the three-phase counter-current chromatogram that obtains fixedly with the splitter (polytetrafluoroethylene (PTFE) twines the helix tube that forms) of the full conventional adverse current chromatogram of the flow pump of 9mL/min, then with the rotating speed antiport adverse current chromatogram main frame of 500r/min; Simultaneously go into mobile phase (being the last phase in the liquid two-phase) with the flow pump of 0.5mL/min; By fixedly under Archimedes spiral power, planetary centrifugal force field and mobile comprehensive function mutually, being retained in the splitter that the solid-liquid two-phase constitutes; Add the back-pressure of 1Mpa at the port of export of adverse current chromatogram, wait to flow when overfall overflows, three alternate balances are set up; 30% fix is retained in the splitter mutually;
4) the rough sample of 8mg root of large-flowered skullcap water extract is dissolved in 4mL is liquid and flows mutually, push, it is entered in the splitter of adverse current chromatogram mutually with flowing via the sample introduction circle;
5) root of large-flowered skullcap water extract is separated purification; Use under the UV-detector 285nm and carry out online detection, and collect each cut, on liquid chromatograph-mass spectrometer,, determine that the 2nd peak is baicalein (purity 92%) each cut and chemical reference substance comparison according to the data (see figure 1) of computer software collection.
The separation and purification of embodiment 2, red sage root fat-soluble compound
Use inverse bonded phase silica gel C18 and n-hexane-alcohol-water (volume ratio is 10-5.5-4.5) system to constitute the three-phase of three-phase counter-current chromatographic system of the present invention, come separation and purification red sage root fat-soluble compound, concrete steps are:
1) according to the volume ratio of 10-5.5-4.5, n-hexane, second alcohol and water are mixed, and in separatory funnel balance 2h, mixed liquor is divided into two-phase up and down, obtains n-hexane-alcohol-water dicyandiamide solution; To be separated up and down;
2) use inverse bonded phase silica gel C18 (carbon 18) (GE Healthcare, average diameter of particles 5 μ m) as solid-state fixedly phase; With dicyandiamide solution Semi-polarity in the step 1) and solid-state fixing mutually close going up mutually as liquid fixedly phase; With inverse bonded phase silica gel C18 and solution mixing mutually down, the addition of C18 is 1g/L with the following ratio of solution mutually; Speed with 60r/min mixes liquid, and inverse bonded phase silica gel C18 is uniformly distributed in the phase solution, obtains the fixedly phase of three-phase counter-current chromatographic system of the present invention, comprising solid-state fixedly phase with liquid fixing mutually;
With dicyandiamide solution Semi-polarity in the step 1) and solid-state fixing close following mobile phase as the three-phase counter-current chromatogram;
3) with step 2) the three-phase counter-current chromatogram that obtains fixedly with the splitter (polytetrafluoroethylene (PTFE) twines the helix tube that forms) of the full conventional adverse current chromatogram of the flow pump of 9mL/min, then with the rotating speed forward running adverse current chromatogram main frame of 600r/min; Simultaneously go into mobile phase (being the last phase in the liquid two-phase) with the flow pump of 0.5mL/min; By fixedly under Archimedes spiral power, planetary centrifugal force field and mobile comprehensive function mutually, being retained in the splitter that the solid-liquid two-phase constitutes; Add the back-pressure of 0.7Mpa at the port of export of adverse current chromatogram, wait to flow when overfall overflows, three alternate balances are set up; 38% fix is retained in the splitter mutually;
4) the fat-soluble rough sample of the 10mg red sage root is dissolved in 3mL is liquid and flows mutually, push, it is entered in the splitter of adverse current chromatogram mutually with flowing via the sample introduction circle;
5) the fat-soluble rough sample of the red sage root is separated purification; Use under the UV-detector 280nm and carry out online detection, and the data (see figure 2) of gathering according to computer software, collect each cut, on liquid chromatograph-mass spectrometer,, determine that the 5th peak is tanshinone IIA (purity 95%) each cut and chemical reference substance comparison.
Claims (3)
1, a kind of method of using three-phase counter-current chromatographic separation and purification natural products, its be the adverse current chromatogram of routine fixing mutually in, adding solid-state separating medium---microballoon is as solid-state fixedly phase, form a piece-rate system that comprises liquid fixedly phase, solid-state fixedly phase and liquid mobile three-phase mutually, the method that adverse current chromatogram method and column chromatography method are combined; Comprise following step:
1) select conventional adverse current chromatogram dicyandiamide solution to flow mutually as the liquid state of three-phase counter-current chromatogram is mutually fixing and liquid, and in separatory funnel balance 2h, two-phase is separated up and down, and is standby;
2) the fixedly phase of preparation three-phase counter-current chromatogram flows mutually with liquid:
The adverse current chromatogram dicyandiamide solution Semi-polarity of the routine described in the step 1) is fixed a mutually close phase as liquid fixedly phase with solid-state; Then with solid-state fixedly be added to above-mentioned liquid state fixing mutually in, the solid-state of adding is 2～6g/L with the fixing ratio mutually of liquid state fixedly, mixes, the suspension that makes is as the fixedly phase of three-phase counter-current chromatogram;
Described solid-state fixing is lipophilic medium or hydrophilic medium mutually;
With the adverse current chromatogram dicyandiamide solution Semi-polarity of the routine described in the step 1) and a solid-state fixing close liquid state as the three-phase counter-current chromatogram phase that flows;
3) set up three alternate balances:
With step 2) obtain contain lipophilic medium fixedly with the splitter of the full conventional adverse current chromatogram of the flow pump of 8～10mL/min, then with the rotating speed forward running adverse current chromatogram main frame of 300～600r/min; Pump into the liquid phase that flows with low flow velocity 0.5～1.5mL/min simultaneously; Add the back-pressure of 0.7Mpa at the port of export of adverse current chromatogram, wait to flow when overfall overflows, three alternate balances are set up; Under Archimedes spiral power, planetary centrifugal force field and mobile comprehensive function mutually, 25～70% fix is retained in the splitter mutually;
Or with step 2) obtain contain hydrophilic media fixedly with the splitter of the full conventional adverse current chromatogram of the flow pump of 8～10mL/min, then with the rotating speed antiport adverse current chromatogram main frame of 300～600r/min; Pump into the liquid phase that flows with low flow velocity 0.5～1.0mL/min simultaneously; Add the back-pressure of 1Mpa at the port of export of adverse current chromatogram, wait to flow when overfall overflows, three alternate balances are set up; Under Archimedes spiral power, planetary centrifugal force field and mobile comprehensive function mutually, 25～60% fix is retained in the splitter mutually;
4) with the natural products sample introduction of purifying to be separated:
With the natural products of purifying to be separated by the concentration of 1～10mg/mL be dissolved in liquid flow mutually after, push the sample introduction circle;
5) separation and purification:
In the natural products sample of purifying to be separated each component liquid state fixing with distribution between liquid state flows mutually in, also adsorption chromatography take place mutually separate with solid-state fixing; Use the online detection cut of UV-detector, and by recorder or computer software record chromatogram, and, obtain the natural products of purifying according to chromatogram collection cut;
The liquid conventional adverse current chromatogram dicyandiamide solution of step 1) is n-hexane-alcohol-water, n-hexane-ethyl acetate-methanol-water, n-hexane-ethyl acetate-methyl alcohol-acetate-water, ethyl acetate-n-butanol-acetonitrile-water, ethyl acetate, alcohol and water, ethyl acetate-ethanol-acetate-water, ethyl acetate-n-butanol-water, n-butanol-water or n-butanol-acetate-water.
2, the method for use three-phase counter-current chromatographic separation and purification natural products as claimed in claim 1 is characterized in that: step 2) described lipophilic medium is the cross-linked copolymer of inverse bonded phase silica gel, polystyrene, polyamide or styrene-divinylbenzene.
3, the method for use three-phase counter-current chromatographic separation and purification natural products as claimed in claim 1 is characterized in that: step 2) described hydrophilic medium is positive bonding phase silica gel, agarose matrix gel or glucan matrix gel.
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| CN108299404B (en) * | 2018-01-08 | 2021-04-06 | 宁波大学 | Method for extracting and separating natural product by liquid-liquid three-phase solvent system and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1135522A (en) * | 1965-09-15 | 1968-12-04 | Elmar Viking Piel | Chromatographic process |
| US4430216A (en) * | 1982-09-09 | 1984-02-07 | The United States Of America As Represented By The Department Of Health And Human Services | High speed preparative countercurrent chromatography with a multiple layer coiled column |
| CN1323776A (en) * | 2001-05-31 | 2001-11-28 | 北京天纯维通生物技术有限公司 | Prepn. of resveratrol and resveratrol glycoside |
| US6337021B1 (en) * | 1994-12-16 | 2002-01-08 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Chiral separation of enantiomers by high-speed countercurrent chromatography |
| US20040241878A1 (en) * | 2002-09-13 | 2004-12-02 | Jorg Thommes | Method of purifying polypeptides by simulated moving bed chromatography |
| CN1634875A (en) * | 2004-11-11 | 2005-07-06 | 中国人民解放军第二军医大学 | Process for rapid preparation of high pure pharmaceutical matters from patrinia villosa juss |
-
2005
- 2005-09-23 CN CNB2005100865159A patent/CN100434135C/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1135522A (en) * | 1965-09-15 | 1968-12-04 | Elmar Viking Piel | Chromatographic process |
| US4430216A (en) * | 1982-09-09 | 1984-02-07 | The United States Of America As Represented By The Department Of Health And Human Services | High speed preparative countercurrent chromatography with a multiple layer coiled column |
| US6337021B1 (en) * | 1994-12-16 | 2002-01-08 | The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services | Chiral separation of enantiomers by high-speed countercurrent chromatography |
| CN1323776A (en) * | 2001-05-31 | 2001-11-28 | 北京天纯维通生物技术有限公司 | Prepn. of resveratrol and resveratrol glycoside |
| US20040241878A1 (en) * | 2002-09-13 | 2004-12-02 | Jorg Thommes | Method of purifying polypeptides by simulated moving bed chromatography |
| CN1634875A (en) * | 2004-11-11 | 2005-07-06 | 中国人民解放军第二军医大学 | Process for rapid preparation of high pure pharmaceutical matters from patrinia villosa juss |
Non-Patent Citations (3)
| Title |
|---|
| Supercritical fluid extraction of aurentiamide acetatefrom Patrinia villosa Juss and subsequent isolation by silica gelandhigh-speed counter-current chromatography. Jinyong Peng et al.Journal of Chromatography A,Vol.1083. 2005 * |
| 高速逆流色谱双水相体系分离蛋白质. 郅文波等.色谱，第23卷第1期. 2005 * |
| 高速逆流色谱技术在生物大分子分离纯化中的应用. 郅文波等.生物技术通讯，第16卷第1期. 2005 * |
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