CN109091911B - Double-column switching cycle chromatographic separation system and method for separating and concentrating target components from raw materials - Google Patents
Double-column switching cycle chromatographic separation system and method for separating and concentrating target components from raw materials Download PDFInfo
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- CN109091911B CN109091911B CN201811024239.7A CN201811024239A CN109091911B CN 109091911 B CN109091911 B CN 109091911B CN 201811024239 A CN201811024239 A CN 201811024239A CN 109091911 B CN109091911 B CN 109091911B
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Abstract
A dual-column switching cycle chromatographic separation system and method for separating and concentrating target components from a feedstock includes first and second chromatographic columns, a valve assembly; the two chromatographic columns are connected in series through a valve component and are switched at regular intervals, the eluent and the modifier respectively flow in from inlets of the two chromatographic columns, the elution capacity of the eluent is stronger than that of the modifier, and the process is as follows: the first chromatographic column and the second chromatographic column are connected in series, eluent flows in from the inlet of the first chromatographic column and modifier flows in from the inlet of the second chromatographic column, and residual liquid flows out from the outlet of the second chromatographic column; the second chromatographic column is connected with the first chromatographic column in series, eluent flows in from the inlet of the second chromatographic column and modifier flows in from the inlet of the first chromatographic column, and residual liquid flows out from the outlet of the first chromatographic column; the first chromatographic column and the second chromatographic column are connected in series, eluent flows in from the inlet of the first chromatographic column and modifier flows in from the inlet of the second chromatographic column, and residual liquid flows out from the outlet of the second chromatographic column; the double columns are switched in a reciprocating manner. Has the advantages of simple structure, few solvent types, convenient post-treatment and low cost.
Description
Technical Field
The invention relates to the technical field of chromatographic separation, in particular to a double-column switching cycle chromatographic separation system and a method for separating and concentrating target components from raw materials.
Background
The chromatographic separation method is a common method for separating a target component from a mixture, and has wide application in many fields such as drug separation, natural product separation and purification, and the like. For some systems which are difficult to separate, such as chiral drug resolution, isomer separation and the like, satisfactory separation cannot be obtained usually by single chromatographic operation, and at the moment, the length of a chromatographic column is most simply and effectively prolonged. However, the column pressure increases due to the elongation of the column, and the requirements for the liquid delivery equipment and the column are increased. At this time, a single column circulation chromatography may be employed, as shown in fig. 1, in which a solute concentration band is introduced into the same column again by an infusion pump after flowing out of the column, which is equivalent to extending the column, but when the concentration band passes through the infusion pump during the circulation process, it causes severe band expansion, thereby breaking the separation obtained in the column; therefore, another two-column switching cycle chromatography has been developed, as shown in fig. 2, in which two chromatographic columns are connected by a six-way valve, and the concentration band moves circularly in a closed loop formed by the two columns by switching the valves: when the concentration spectral band leaves the left post and gets into the right post, the valve switches, and the concentration spectral band will pass through the right post and reentry the left post this moment, when the concentration spectral band all leaves the right post and gets into the left post, switches the post again, and the circulation is reciprocal like this, guarantees that the concentration spectral band is two post inner loop removal about all the time, and need not to flow through the transfer pump.
The separation will improve with increasing number of cycles. However, regardless of whether a single column cycle or a two-column switching cycle is employed, the separation component concentration band inevitably broadens during separation, so that when overloaded for preparative separation, the band front edge of the weakly retained component will soon catch up with the band back edge of the strongly retained component, thereby causing the separation to fail.
Peak cutting techniques can also be used to cut the leading edge of the weakly retained component band and the trailing edge of the strongly retained component band that have achieved separation into the system in each cycle, while the overlapping portions of the separated component bands are cycled through the system. This avoids the weak retained component band front following the strong retained component band back, but due to band spreading, the concentration of the acceptable fraction collected in each cycle will be increasingly dilute, thereby increasing the cost of recovering solute from the subsequent vaporized fraction. In some separation processes, such as natural product separation and purification, the recovery operation may determine the cost of the whole separation process due to the low content of the target component.
Therefore, in order to fully exploit the advantages of the cyclic chromatography, consideration must be given to how the band of the concentration of the separated component can be effectively prevented from spreading during the separation process.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the double-column switching cycle chromatographic separation system which has the advantages of simple structure, less solvent types, convenient post-treatment and low cost and is used for separating and concentrating target components from raw materials.
In order to solve the above technical problems, the technical solution provided by the present invention is realized as follows: a dual column switching cycle chromatographic separation system for separating and concentrating a target component from a feed material, the system comprising: a first chromatographic column and a second chromatographic column, a valve assembly; the first chromatographic column and the second chromatographic column are always connected in series through a valve assembly, the series connection mode of the first chromatographic column and the second chromatographic column is switched at regular intervals, an eluant and a modifier used by the system respectively flow into the system from inlets of the two chromatographic columns, and the elution capacity of the eluant is stronger than that of the modifier; the specific switching process is as follows: firstly, a first chromatographic column and a second chromatographic column are connected in series, eluent flows into a system from a first chromatographic column inlet and modifier flows into the system from a second chromatographic column inlet, and residual liquid flows out from a second chromatographic column outlet; after a certain time, the system is switched, so that the second chromatographic column and the first chromatographic column are connected in series, eluent flows into the system from the inlet of the second chromatographic column and modifier flows into the system from the inlet of the first chromatographic column, and residual liquid flows out from the outlet of the first chromatographic column; after a certain time, the system is switched again, so that the first chromatographic column and the second chromatographic column are connected in series, the eluent flows into the system from the inlet of the first chromatographic column and the modifier flows into the system from the inlet of the second chromatographic column respectively, and the residual liquid flows out from the outlet of the second chromatographic column; circularly and repeatedly switching the double columns in such a way to change the positions of the inlets of the eluent and the modifier and the position of the outlet of the residual liquid; finally, the separation and concentration of the target components are realized.
Preferably, in order to make the elution capacity of the eluent stronger than the elution capacity of the modifying agent, the eluent can be heated and the modifying agent can be cooled to ensure that the temperature of the eluent is higher than that of the modifying agent, so that the elution capacity of the eluent is stronger than that of the modifying agent.
Preferably, in order to make the elution ability of the eluent stronger than that of the modifier, a method may be employed in which the volume percentage of the solvent having a strong elution ability in the eluent is higher than that of the solvent having a strong elution ability in the modifier.
Or simultaneously heating the eluent and cooling the modifier and the solvent with strong elution capacity in the eluent by volume percentage higher than that of the solvent with strong elution capacity in the modifier, so as to realize that the elution capacity of the eluent is stronger than that of the modifier.
The invention also provides a method for separating and concentrating target components by using the double-column switching cycle chromatographic separation system, which comprises the following specific steps:
(1) feeding: dissolving the raw material to be separated into any solvent capable of dissolving the raw material to prepare a raw material solution; the initial state of the double-column switching cycle chromatographic separation system is that a first chromatographic column and a second chromatographic column are connected in series, then raw material liquid is added from an inlet of the first chromatographic column, and a target object is adsorbed in the system; in the feeding stage, the two pumps for conveying the eluent and the modifier are not operated, and the valve component is not switched.
(2) And (3) purification: the state of the double-column switching system is still that the first chromatographic column and the second chromatographic column are connected in series, then eluent and modifier respectively flow into the system from the inlet of the first chromatographic column and the inlet of the second chromatographic column, and residual liquid flows out from the outlet of the second chromatographic column; after a certain time, the system is switched, the second chromatographic column is connected with the first chromatographic column in series, the eluent and the modifier respectively flow into the system from the inlet of the second chromatographic column and the inlet of the first chromatographic column, and the residual liquid flows out from the outlet of the first chromatographic column; after a certain time, the system is switched again, the first chromatographic column and the second chromatographic column are connected in series, the eluent and the modifier respectively flow into the system from the inlet of the first chromatographic column and the inlet of the second chromatographic column, and the residual liquid flows out from the outlet of the second chromatographic column; the double columns are switched repeatedly in such a cycle to change the inlet positions of the eluent and the modifier, and finally the purified target product is obtained.
The switching time of the positions of the eluent and the modifier inlet and the effluent outlet at regular intervals (after a certain time) is set according to specific chromatographic columns and separating substances and is generally switched once within 2-20 min.
The system of the present invention stops the feeding when the target component starts to flow out of the first chromatography column during the feeding, and the corresponding feeding amount is the optimum feeding amount. During feeding, the elution pump and the modification pump are not operated, and the valve component is not switched.
In the system switching described in the present invention, the time intervals of each switching may be equal or unequal, that is, the time intervals of two adjacent switching may be equal or unequal.
The first chromatographic column and the second chromatographic column or the second chromatographic column and the first chromatographic column are connected in series, and are defined from the flowing direction of a medium flowing in the system: the fluid flows from the first chromatographic column to the second chromatographic column, and the first chromatographic column is connected with the second chromatographic column in series; and (3) enabling the fluid to flow from the second chromatographic column to the first chromatographic column, namely connecting the second chromatographic column with the first chromatographic column in series. The two serial states are the result of valve switching, and the circular chromatographic operation is switched in the two states to ensure that the whole or overlapped part of the spectral bands circularly reciprocate in the chromatographic column, and the function of each chromatographic column is separated.
The technical principle of the invention is as follows: in dual column switching cycle chromatography, the separation component concentration bands move cyclically within a closed loop formed by two columns, and if the mobile phase in the upstream column elutes more strongly than the mobile phase in the downstream column during band migration, the migration rate of the trailing edge of each separation component concentration band is faster than the migration rate of the leading edge of its concentration band, thus creating a narrowing effect on the concentration band to overcome and counteract the expansion of the concentration band.
The present invention is based on the recognition that the elution capacity of the mobile phase inside the upstream column is stronger than that of the mobile phase inside the downstream column (i.e. the elution capacity of the eluent is stronger than that of the modifier) by the following three measures:
(1) the volume percentage content of the solvent with strong elution capability in the eluent is higher than that of the solvent with strong elution capability in the modifier;
(2) the temperature of the eluent is higher than that of the modifier;
(3) simultaneously, the volume percentage content of the solvent with strong elution capability in the eluent is higher than that of the solvent with strong elution capability in the modifier, and the temperature of the eluent is higher than that of the modifier.
The method of the invention also comprises a final target component recovery step, and can be realized by adopting the following two methods according to the requirement on the purity of the target product:
during the purification process, the concentrations in the system effluent and the feed solution are monitored to ensure that the ratio of the concentrations of the target component to the non-target component in the system effluent (i.e., the concentration of the target component divided by the concentration of the non-target component) is less than the ratio of the concentrations of the target component to the non-target component in the feed solution; with the progress of the purification process, the content ratio of the target component to the non-target component in the system (namely the content of the target component is divided by the content of the non-target component) is gradually increased, when the content ratio of the target component to the non-target component in the system reaches the requirement (the target component to the non-target component in the effluent is detected, the volume of the effluent can be measured, the mass of the target component and the non-target component flowing out of the system in the purification process can be obtained, and the mass of the target component and the non-target component added into the system in the feeding stage is determined, so that the mass of each component flowing out of the system in the purification stage is subtracted from the mass of each component added into the system in the feeding stage, the mass and the content of each component in the system can be obtained), the purification operation is stopped, and the first chromatographic column and the second chromatographic column are eluted to recover the target;
or
Monitoring the concentration of target and non-target components in the effluent of the system and the volume of the effluent during the purification process to ensure that the mass of target and non-target components in the effluent is less than the mass of target and non-target components added to the system during the feed phase; so as the purification process proceeds, the difference in spatial distribution of the concentration bands of the target component and the non-target component inside the system increases gradually; when the separation of the target component and the non-target component concentration band meets the requirement, stopping the purification operation; at the moment, if the first chromatographic column and the second chromatographic column are connected in series, eluting the first chromatographic column and the second chromatographic column, and sequentially collecting non-target components and target components at the outlet of the second chromatographic column; and if the second chromatographic column and the first chromatographic column are connected in series, eluting the second chromatographic column and the first chromatographic column, and sequentially collecting the non-target component and the target component at the outlet of the first chromatographic column.
When the second recovery method of the present invention is adopted, a detector can be arranged at the connection position of the two columns to monitor the separation of the target component and the non-target component concentration spectral bands, and the qualitative judgment of whether the separation requirement is met is helped, for example, the two spectral bands are basically separated and the overlapping part is not too much.
The invention has the advantages and beneficial effects that: the invention provides a method for synchronously separating and concentrating a target component from a raw material by adopting double-column switching cycle chromatography, which can improve the purity of the target component and increase the concentration of the target component in a product solution, thereby reducing the burden of recovering the target component from the product solution. The double-column switching cycle chromatographic separation system provided by the invention has the advantages of simple structure, convenience in operation, simple treatment after separation and purification, and reduction of separation and purification costs.
Drawings
FIG. 1 is a schematic diagram of a single column cycle chromatographic separation system of the prior art.
FIG. 2 is a schematic diagram of a prior art dual column switching cycle chromatographic separation system.
FIG. 3 is a schematic diagram of a first chromatographic column and a second chromatographic column of a dual-column switching cycle chromatographic separation system according to the present invention, which are connected in series.
FIG. 4 is a schematic diagram of a two-column switching cycle chromatographic separation system according to the present invention, in which the second chromatographic column is connected in series with the first chromatographic column.
Detailed Description
The present invention is described in further detail below by way of examples, but the present invention is not limited to only the following examples.
The system is a dual-column switching cycle chromatographic separation system for separating and concentrating target components from raw materials, which is described and protected by the invention; the eluent can be heated and the modifier can be cooled, namely the eluent is ensured to have the temperature higher than that of the modifier, so that the elution capacity of the eluent is stronger than that of the modifier, the eluent is not heated or cooled intentionally, and other modes which can realize the elution capacity of the eluent is stronger than that of the modifier are all suitable for the invention; the columns of the present invention are the two chromatographic columns mentioned in the present system.
The technical scheme of the invention is explained in detail below by taking the separation and purification of orlistat as an example. Orlistat starting material was purchased from biotechnology limited, north Hebei Haidede. On a carbon octadecyl bonded silica gel column, acetonitrile/water (88/12, v/v) is used as a mobile phase, and a plurality of front impurity peaks and rear impurity peaks are arranged before and after a main peak of orlistat, wherein the peak area percentage of each impurity is higher than 0.1%. The invention provides a double-column switching cycle chromatographic separation method for synchronously separating and concentrating target components from raw materials, which is used for separating and concentrating orlistat serving as a main component from orlistat raw materials and ensuring that the peak area percentage of each impurity is lower than 0.1 percent. The switching time of the eluent and the modifier inlet and the effluent outlet at regular intervals is set according to specific chromatographic columns and separating substances and is switched once in 2-20 min.
1. Double-column switching circulating chromatographic separation system
Shown in figures 3-4: the invention relates to a double-column switching cycle chromatographic separation system, which comprises: a first chromatographic column 1, a second chromatographic column 2 and a valve assembly 3 (specifically, two groups of six-way valves 3); the first chromatographic column and the second chromatographic column are always connected in series through two groups of six-way valves, the series connection mode of the first chromatographic column and the second chromatographic column is switched at regular intervals, eluent and modifier used by the system respectively flow into the system from inlets of the two chromatographic columns (the first chromatographic column and the second chromatographic column), the elution capacity of the eluent is stronger than that of the modifier, and raw material liquid of the system can enter the system from the inlet of the first chromatographic column or can be added into the system from the inlet of the second chromatographic column (the efficiency is slightly lower); the specific switching process is as follows: firstly, as shown in fig. 3, a first chromatographic column and a second chromatographic column are connected in series, and the raw material liquid and the eluent respectively flow into the system from the inlet of the first chromatographic column and the modifier respectively from the inlet of the second chromatographic column; after a certain time, the system is switched, specifically as shown in fig. 4, the second chromatographic column and the first chromatographic column are connected in series, and the eluent flows into the system from the inlet of the second chromatographic column and the modifier flows into the system from the inlet of the first chromatographic column respectively; after a certain time, the system is switched again, specifically as shown in fig. 3, the first chromatographic column is connected with the second chromatographic column in series again, and the eluent flows into the system from the inlet of the first chromatographic column and the modifying agent flows into the system from the inlet of the second chromatographic column respectively; the double columns are switched in a reciprocating mode in such a cycle, so that the positions of inlets of the eluent and the modifier are changed;
the system of the present invention, during the feeding, stops the feeding if the raw material liquid is fed from the first chromatographic column when the target component starts to flow out from the first chromatographic column; stopping the feeding if the feed solution is fed from the second chromatography column when the target component begins to flow from the second chromatography column; the corresponding feed rate is then the optimum feed rate. During feeding, the elution pump and the modification pump are not operated, and the valve component is not switched.
In the system: the raw material liquid conveying pipeline and the eluent (eluent) conveying pipeline are connected through a three-way joint, and a plunger pump 4 is arranged on the pipelines of the raw material liquid, the eluent and the modifier to provide power for the circulation flow of the medium in the system.
The dual-column switching cycle chromatographic separation system of the present embodiment mainly comprises three plunger pumps 4 for delivering a raw material liquid, an eluent and a modifier, and two chromatographic columns (inner diameter 1cm, length 20cm) filled with C18 silica gel; the eluent is ethanol/water (88/12, v/v), and the water content of the modifying agent is higher than that of the eluent, so the elution capacity of the eluent is stronger than that of the modifying agent. Orlistat raw material was dissolved in ethanol/water (86/14, v/v) to make a raw material solution with a total concentration of 20 mg/mL.
2. Inspection of finished product
And analyzing the purity of the product by high performance liquid chromatography. The high performance liquid chromatography system comprises: a Dalian Eitide 1201 pump, a Phenomenex Knitex XB-18 column (4.6X 150mm, 5 μm), a Dalian Eitide 1201 detector; liquid chromatography conditions: the mobile phase is acetonitrile/water (volume ratio 90/10), the flow rate is 1.0mL/min, the detection wavelength is 210nm, and the column temperature is 30 ℃.
Example 1 separation and concentration of orlistat by a double column switching cycle chromatographic separation System
The separation system is first shown in fig. 3: the first chromatographic column and the second chromatographic column are connected in series, the raw material liquid is added into the system from the inlet of the first chromatographic column at room temperature, and when the volume of the added raw material liquid is 60mL, the separated components just flow out of the first chromatographic column, and the feeding is stopped; then, the plunger pump on the eluent line and the plunger pump on the modifier line were simultaneously started, and eluent and modifier (water) were added to the system from the first chromatography column inlet and the second chromatography column inlet at a flow rate of 4mL/min and 0.2mL/min, respectively. After 10min, the two six-way valves were switched simultaneously, as shown in fig. 4: the second chromatographic column is connected with the first chromatographic column in series, and eluent and the modifying agent respectively flow into the system from inlets of the second chromatographic column and the first chromatographic column at the flow rates of 4m/min and 0.2 mL/min; the above steps are repeated in cycles, and the switching is performed once every 10min for 5 times. In the separation and purification process, the effluent liquid of the system does not contain impurities and orlistat. Then, the valve switching was stopped, the dressing pump was turned off, the elution pump was turned on alone, the first and second chromatography columns were eluted at a flow rate of 5mL/min, and three fractions were collected in the second chromatography column tail section: fraction 1 mainly contains the precursor impurity and a small amount of orlistat, and the area of the precursor impurity peak is 20 times that of the precursor impurity peak in the raw material liquid; the fraction 2 contains orlistat, does not contain pre-impurity and post-impurity, has the concentration of 29mg/mL, is higher than the concentration of orlistat in a raw material solution, and has the yield of orlistat of 92 percent; fraction 3 mainly contains late impurities and orlistat with very dilute concentration, wherein the peak area of the late impurities is 13 times that of the late impurities in the raw material liquid.
Example 2 separation and concentration of orlistat Using a Dual column switching cycle chromatographic separation System
The separation system is first shown in fig. 3: the first chromatographic column and the second chromatographic column are connected in series, the raw material liquid is added into the system from the inlet of the first chromatographic column at room temperature, and when the volume of the added raw material liquid is 60mL, the separated components just flow out of the first chromatographic column, and the feeding is stopped; then, the elution pump and the modification pump were simultaneously started, and an eluent and a modifier (ethanol/water: 10/90(v/v)) were added to the system at a flow rate of 4mL/min and 1.0mL/min from the first chromatography column inlet and the second chromatography column inlet, respectively. When the eluent and the modifier are delivered, the eluent is heated to 45 ℃ and the modifier is cooled to 5 ℃. After 10min, the two six-way valves were switched simultaneously, as shown in fig. 4: the second chromatographic column is connected with the first chromatographic column in series, and eluent and the modifier respectively flow into the system from inlets of the second chromatographic column and the first chromatographic column at the flow rates of 3.5m/min and 1.0 mL/min; the above steps are repeated in cycles, and the switching is performed once every 10min for 6 times. In the separation and purification process, the effluent liquid of the system does not contain impurities and orlistat. Then, the valve switching was stopped, the dressing pump was turned off, the elution pump was turned on alone, the first and second chromatography columns were eluted at a flow rate of 5mL/min, and three fractions were collected in the second chromatography column tail section: fraction 1 mainly contains the precursor impurity and a small amount of orlistat, and the peak area of the precursor impurity is 25 times of that of the precursor impurity in the raw material liquid; fraction 2 contains orlistat, does not contain pre-impurity and post-impurity, and has a concentration of 32mg/mL, which is higher than the concentration of orlistat in the raw material solution; fraction 3 mainly contains late impurities and orlistat with very dilute concentration, wherein the peak area of the late impurities is 15 times that of the late impurities in the raw material liquid.
Through the embodiments of the present system described above, it can be seen that the separation system of the present invention can increase the concentration of the target component in the product solution while improving the purity of the target component, thereby reducing the burden of recovering the target component from the product solution. The double-column switching cycle chromatographic separation system provided by the invention has the advantages of simple structure, convenience in operation, simple treatment after separation and purification, and reduction of separation and purification costs.
The above examples are illustrative and further illustrative of the present invention and are not intended to be limiting, and any modification made within the spirit and scope of the present invention will fall within the scope of the present invention.
Claims (8)
1. A method for separating and concentrating target components from raw materials by using a dual-column switching cycle chromatographic separation system is characterized by comprising the following steps of: the system comprises: a first chromatographic column and a second chromatographic column, a valve assembly; the first chromatographic column and the second chromatographic column are always connected in series through a valve assembly, the series connection mode of the first chromatographic column and the second chromatographic column is switched at regular intervals, an eluant and a modifier used by the system respectively flow into the system from inlets of the two chromatographic columns, and the elution capacity of the eluant is stronger than that of the modifier; the specific switching process is as follows: firstly, a first chromatographic column and a second chromatographic column are connected in series, and the eluent flows into the system from the inlet of the first chromatographic column and the modifier flows into the system from the inlet of the second chromatographic column respectively; after a certain time, the system is switched, so that the second chromatographic column and the first chromatographic column are connected in series, and eluent flows into the system from the inlet of the second chromatographic column and the modifier flows into the system from the inlet of the first chromatographic column respectively; after a certain time, the system is switched again, so that the first chromatographic column and the second chromatographic column are connected in series, and the eluent flows into the system from the inlet of the first chromatographic column and the modifier flows into the system from the inlet of the second chromatographic column respectively; the double columns are switched in a reciprocating mode in such a cycle, so that the positions of inlets of the eluent and the modifier are changed; the time intervals of the system switching are not necessarily equal;
the specific separation steps include:
(1) feeding: dissolving the raw material to be separated into any solvent capable of dissolving the raw material to prepare a raw material solution; the initial state of the double-column switching cycle chromatographic separation system is that a first chromatographic column and a second chromatographic column are connected in series, then raw material liquid is added from an inlet of the first chromatographic column, and a target object is adsorbed in the system;
and (3) purification: the state of the double-column switching system is still that the first chromatographic column and the second chromatographic column are connected in series, then eluent and modifier respectively flow into the system from the inlet of the first chromatographic column and the inlet of the second chromatographic column, and residual liquid flows out from the outlet of the second chromatographic column; after a certain time, the system is switched, the second chromatographic column is connected with the first chromatographic column in series, the eluent and the modifier respectively flow into the system from the inlet of the second chromatographic column and the inlet of the first chromatographic column, and the residual liquid flows out from the outlet of the first chromatographic column; after a certain time, the system is switched again, the first chromatographic column and the second chromatographic column are connected in series, the eluent and the modifier respectively flow into the system from the inlet of the first chromatographic column and the inlet of the second chromatographic column, and the residual liquid flows out from the outlet of the second chromatographic column; the double columns are switched repeatedly in such a cycle to change the inlet positions of the eluent and the modifier, and finally the purified target product is obtained.
2. The method of claim 1 for separating and concentrating a target component from a feedstock using a dual column switching cycle chromatographic separation system, wherein: the elution capacity of the eluent is stronger than that of the modifier, and specifically comprises the following steps: heating the eluent and cooling the modifying agent to achieve a stronger elution capacity of the eluent than the modifying agent.
3. The method of claim 1 for separating and concentrating a target component from a feedstock using a dual column switching cycle chromatographic separation system, wherein: the elution capacity of the eluent is stronger than that of the modifier, and specifically comprises the following steps: the volume percentage content of the solvent with strong elution capability in the eluent is higher than that of the solvent with strong elution capability in the modifier.
4. The method of claim 1 for separating and concentrating a target component from a feedstock using a dual column switching cycle chromatographic separation system, wherein: the elution capacity of the eluent is stronger than that of the modifier, and specifically comprises the following steps: heating eluent and cooling modifier, and controlling the volume percentage content of the solvent with strong elution capacity in the eluent to be higher than that of the solvent with strong elution capacity in the modifier.
5. The method of claim 4 for separating and concentrating a target component from a feedstock using a dual column switching cycle chromatographic separation system, wherein: the time period, i.e. the switching time of the eluent and modifier inlet and effluent outlet positions, is set according to the specific chromatographic column and separation material.
6. The method of claim 4 for separating and concentrating a target component from a feedstock using a dual column switching cycle chromatographic separation system, wherein: the method also comprises a final target component recovery step, which specifically comprises the following steps: during the purification process, monitoring the concentrations of the system effluent and the raw material liquid to ensure that the concentration ratio of the target component to the non-target component in the system effluent is lower than the concentration ratio of the target component to the non-target component in the raw material liquid; and when the content ratio of the target component to the non-target component in the system reaches the requirement, stopping the purification operation, and eluting the first chromatographic column and the second chromatographic column to recover the target component.
7. The method of claim 4 for separating and concentrating a target component from a feedstock using a dual column switching cycle chromatographic separation system, wherein: the method also comprises a final target component recovery step, which specifically comprises the following steps: monitoring the concentration of target and non-target components in the effluent of the system and the volume of the effluent during the purification process to ensure that the mass of target and non-target components in the effluent is less than the mass of target and non-target components added to the system during the feed phase; as the purification process proceeds, the difference in spatial distribution of the concentration bands of the target component and the non-target component inside the system increases progressively; when the separation of the target component and the non-target component concentration band meets the requirement, stopping the purification operation; at the moment, if the first chromatographic column and the second chromatographic column are connected in series, eluting the first chromatographic column and the second chromatographic column, and sequentially collecting non-target components and target components at the outlet of the second chromatographic column; and if the second chromatographic column and the first chromatographic column are connected in series, eluting the second chromatographic column and the first chromatographic column, and sequentially collecting the non-target component and the target component at the outlet of the first chromatographic column.
8. The method of claim 7 for separating and concentrating a target component from a feedstock using a dual column switching cycle chromatographic separation system, wherein: in monitoring the concentration of target and non-target components in the effluent of the system and the volume of the effluent, a detector is placed in the connection between the two chromatography columns to monitor the separation of the target and non-target component concentration bands.
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