Single-column and double-column alternating cyclic chromatography system and method for separation and purification

Abstract

Disclosed in the present invention are a single-column and double-column alternating cyclic chromatography system and method for separation and purification. The system comprises a first chromatographic column, a second chromatographic column, a first detector, a second detector, a first control valve, a second control valve, a first multi-way valve, a second multi-way valve, a third multi-way valve, a fourth multi-way valve, a fifth multi-way valve, and a sixth multi-way valve. The first control valve, the first multi-way valve, the first chromatographic column, the first detector, and the third multi-way valve are connected in sequence. The second control valve, the second multi-way valve, the second chromatographic column, the second detector, and the fourth multi-way valve are connected in sequence. The first multi-way valve is connected to the second control valve. The second multi-way valve is also connected to the first control valve. The second multi-way valve is connected to the third multi-way valve, the fifth multi-way valve, and the sixth multi-way valve. The fifth multi-way valve and the sixth multi-way valve are also connected to the first multi-way valve. The fourth multi-way valve is also connected to the first multi-way valve, the fifth multi-way valve, and the sixth multi-way valve. The present invention achieves a simple device structure, high separation efficiency, and a high yield.

Description

Single and dual column alternating cyclic chromatography systems and methods for separation and purification Technical Field

[0001] This application relates to the field of chromatographic detection technology, and in particular to a single-column and dual-column alternating cyclic chromatographic system and method for separation and purification. Background Technology

[0002] In the field of chromatographic detection, traditional single-column chromatography is typically an intermittent operation, requiring continuous steps of feeding, elution, and collection. This process is cumbersome, time-consuming, and requires a large amount of eluent, resulting in high production costs. Furthermore, traditional single-column chromatography often yields a small number of qualified samples after a single purification, necessitating secondary concentration or dilution of the sample and impurities before multiple purification cycles. Existing simulated moving bed (SMB) systems utilize specialized equipment and automated control technology to achieve countercurrent flow between the stationary and mobile phases. This allows for more thorough contact between the separated substances and the stationary phase, effectively separating different components in a mixture. Higher purity samples are obtained with less eluent consumption, and the system can be automated for continuous operation, reducing labor costs. However, traditional simulated moving bed (SMB) systems usually require multiple columns and more complex piping and valve systems to function as a simulated moving bed. This results in complex equipment structures, large footprints, high investment costs, and difficulties in cleaning and maintenance. Summary of the Invention

[0003] Therefore, it is necessary to provide a single-column / dual-column alternating cyclic chromatography system for separation and purification. The single-column / dual-column alternating cyclic chromatography system of the present invention is compact, simple in structure, requires fewer chromatographic columns, is easy to operate, and produces high purity and high yield of the separated products, enabling automated and continuous separation and purification.

[0004] One embodiment of this application provides a single-column and dual-column alternating cyclic chromatography system for separation and purification.

[0005] A single / dual-column alternating cyclic chromatography system for separation and purification includes a first column, a second column, a first detector, a second detector, a first control valve, a second control valve, a first multi-port valve, a second multi-port valve, a third multi-port valve, a fourth multi-port valve, a fifth multi-port valve, and a sixth multi-port valve. The first control valve, the first multi-port valve, the first column, the first detector, and the third multi-port valve are connected sequentially. The second control valve, the second multi-port valve, the second column, the second detector, and the fourth multi-port valve are also connected sequentially. The first multi-port valve is further connected to the second control valve, and the second multi-port valve is also connected to the first control valve. The second multi-port valve is simultaneously connected to the third, fifth, and sixth multi-port valves. The fifth and sixth multi-port valves are also simultaneously connected to the first multi-port valve, and the fourth multi-port valve is also simultaneously connected to the first, fifth, and sixth multi-port valves.

[0006] In some embodiments, the first control valve, the second control valve, the first multi-way valve, the second multi-way valve, the third multi-way valve, the fourth multi-way valve, the fifth multi-way valve, and the sixth multi-way valve are each independently selected from four-way valves.

[0007] In some embodiments, the single-column alternating cyclic chromatography system for separation and purification further includes a first drive pump, wherein one port of the first multi-way valve is connected in parallel with one port of the second multi-way valve and then connected to the first control valve via the first drive pump.

[0008] In some embodiments, the single / dual column alternating cyclic chromatography system for separation and purification further includes a second drive pump, wherein one port of the first multi-way valve is connected in parallel with one port of the second multi-way valve and then connected to the second control valve via the second drive pump.

[0009] In some embodiments, the single / dual column alternating cyclic chromatography system for separation and purification further includes a third drive pump, and one of the valve ports of the fourth multi-port valve, one of the valve ports of the fifth multi-port valve, and one of the valve ports of the sixth multi-port valve are connected in parallel to the first multi-port valve via the third drive pump.

[0010] In some embodiments, the single / dual column alternating cyclic chromatography system for separation and purification further includes a fourth drive pump connected to one of the ports of the fifth multi-port valve for pumping in diluent.

[0011] In some embodiments, the single / dual column alternating cyclic chromatography system for separation and purification further includes a fifth drive pump connected to one of the ports of the sixth multi-port valve for pumping out overflow solvent.

[0012] In some embodiments, the first detector includes one or more of an ultraviolet detector, an evaporative light detector, and a differential refractive index detector.

[0013] In some embodiments, the second detector includes one or more of an ultraviolet detector, an evaporative light detector, and a differential refractive index detector.

[0014] In some embodiments, the first multi-port valve includes at least a first valve port connected to a first control valve, a second valve port connected to a second control valve, a third valve port connected to a first chromatographic column, and a fourth valve port connected to a third drive pump.

[0015] The second multi-way valve includes at least a first valve port connected to the first control valve, a second valve port connected to the second control valve, a third valve port connected to the second chromatographic column, and a fourth valve port simultaneously connected to the fifth multi-way valve and the sixth multi-way valve;

[0016] The third multi-way valve includes at least a first valve port connected to the first detector and used to discharge pre-impurities, a second valve port connected to both the second and fifth multi-way valves, a third valve port used to discharge post-impurities, and a fourth valve port used to discharge separated samples.

[0017] The fourth multi-way valve includes at least a first valve port connected to the second detector and used for discharging impurities, a second valve port used for discharging impurities, a third valve port used for discharging separated samples, and a second valve port connected simultaneously to the third drive pump, the fifth multi-way valve, and the sixth multi-way valve.

[0018] The fifth multi-way valve includes at least a first valve port, a second valve port connected to the second multi-way valve, a third valve port for pumping in diluent, and a fourth valve port connected to both the first and fourth multi-way valves.

[0019] The sixth multi-way valve includes at least a first valve port, a second valve port connected to the second multi-way valve, a third valve port for discharging overflow solvent, and a fourth valve port connected to both the first and fourth multi-way valves.

[0020] One embodiment of this application also provides a single-column and dual-column alternating cyclic chromatography method for separation and purification.

[0021] A single-column / dual-column alternating cyclic chromatography method for separation and purification, using the aforementioned single-column / dual-column alternating cyclic chromatography system for separation and purification, includes the following steps:

[0022] In the first cycle, the multi-component mixture (G) is saturated and loaded onto the first chromatographic column. The column is then eluted, and the multi-component mixture (G) is developed and sequentially separated into a pre-impurity (a), a pre-impurity and sample mixture (b), a small amount of acceptable sample (c), a post-impurity and sample mixture (d), and a post-impurity section (e). Under elution with the mobile phase, the pre-impurity (a) is discharged first through the third multi-port valve. The pre-impurity and sample mixture (b) is transferred to the second chromatographic column, and the small amount of acceptable sample (c) is transferred to... In the second column, the impurity-sample mixture (d) is transferred to the second column. The impurity-sample mixture (b), a small amount of qualified sample, and the impurity-sample mixture (d) together form the internal circulation sample (f). During the transfer of the internal circulation sample (f), the entire system is controlled in a closed loop state, without the need to add additional elution solvent. At this stage, diluent is pumped in for online dilution according to the application scenario of the sample. The first control valve switches to regenerator to discharge the impurity (e) from the first column. The first control valve switches to elution solvent to equilibrate the first column, and the first cycle ends.

[0023] The second cycle begins by transferring the internal circulation sample (f) from the second column to the first column through the inlet of the first column used to receive the internal circulation sample (f). During the transfer of the internal circulation sample (f), the entire system is kept in a closed loop state without the need for additional elution solvent. At this time, diluent is pumped in for online dilution according to the sample application scenario. The first control valve is switched to regenerant, and the first drive pump is turned on to discharge the impurities (e) from the second column. The first control valve is switched to elution solvent, and the first drive pump is turned on to equilibrate the second column. The second cycle ends.

[0024] And repeat the first and second cycles until the inner circulation sample (f) meets the quality requirements.

[0025] In some embodiments, the impurities (a) or impurities (e) at the effluent of the first chromatographic column and the second chromatographic column are collected and stored respectively.

[0026] In some embodiments, the mobile phase used by the first drive pump includes, but is not limited to, eluent, regeneration solution, or equilibration solution.

[0027] In some embodiments, after repeating the first and second cycles at least once, the first chromatographic column and / or the second chromatographic column are regenerated or equilibrated by pumping regeneration solution and equilibration solution into the first drive pump.

[0028] The aforementioned single-column and dual-column alternating cyclic chromatography system for separation and purification is compact, simple in structure, requires fewer chromatographic columns, is easy to operate, and produces high purity and high yield of the separated products, enabling automated and continuous separation and purification.

[0029] This invention achieves continuous and automated cyclic operation of dual-column chromatography by reducing the number of chromatographic columns and utilizing the countercurrent operation of a simulated moving bed (SMB). Its main advantages are that it uses two chromatographic columns to achieve the functions of simulated moving bed countercurrent operation and continuous operation, enabling true continuous separation and purification of multi-component mixtures with lower investment costs. It offers advantages such as simpler equipment structure, flexible and convenient operation, higher separation efficiency and yield, automated continuous operation, saving time and labor costs, low investment costs, easy maintenance and cleaning, and significantly reduced eluent usage. Attached Figure Description

[0030] To more clearly illustrate the technical solutions in the embodiments of this application, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application, and those skilled in the art can obtain other drawings based on these drawings without creative effort.

[0031] To gain a more complete understanding of this application and its beneficial effects, the following description will be provided in conjunction with the accompanying drawings. In the following description, the same reference numerals denote the same parts.

[0032] Figure 1 is a schematic diagram of the control of a single-column and dual-column alternating cyclic chromatography system for separation and purification according to an embodiment of the present invention;

[0033] Figure 2 is a liquid chromatography chromatogram of a certain oil preparation according to an embodiment of the present invention;

[0034] Figure 3 shows the detection spectrum of the oil after separation and purification using the single and dual column alternating cyclic chromatography system described in an embodiment of the present invention.

[0035] Explanation of reference numerals in the attached figures

[0036] 10. A single / dual column alternating cyclic chromatography system for separation and purification; 110. First column; 120. Second column; 210. First detector; 220. Second detector; 310. First control valve; 320. Second control valve; 330. First multi-port valve; 340. Second multi-port valve; 350. Third multi-port valve; 360. Fourth multi-port valve; 370. Fifth multi-port valve; 380. Sixth multi-port valve; 410. First drive pump; 420. Second drive pump; 430. Third drive pump; 440. Fourth drive pump; 450. Fifth drive pump. Detailed Implementation

[0037] To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Many specific details are set forth in the following description to provide a thorough understanding of the present invention. However, the present invention can be practiced in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0038] In the description of this invention, it should be understood that the terms "center," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," and "circumferential" indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.

[0039] In this invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," "linking," and "fixing," etc., should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral part; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to the internal communication of two components or the interaction between two components, unless otherwise explicitly limited. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0040] In this invention, unless otherwise explicitly specified and limited, "above" or "below" the second feature can mean that the first feature is in direct contact with the second feature, or that the first feature is in indirect contact with the second feature through an intermediate medium. Furthermore, "above," "over," and "on top" of the second feature can mean that the first feature is directly above or diagonally above the second feature, or simply that the first feature is at a higher horizontal level than the second feature. "Below," "below," and "under" the second feature can mean that the first feature is directly below or diagonally below the second feature, or simply that the first feature is at a lower horizontal level than the second feature.

[0041] In the description of this invention, "several" means one or more, "multiple" means two or more, "greater than," "less than," and "exceeding" are understood to exclude the stated number, while "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0042] In this document, "optionally," "optionally," and "optional" mean that something is optional, that is, it is selected from either "with" or "without." If multiple "options" appear in a technical solution, unless otherwise specified and there are no contradictions or mutual constraints, each "option" is independent. In this application, descriptions such as "optionally contains" and "optionally includes" indicate "contains or does not contain."

[0043] In this invention, unless otherwise stated, the sum of the parts of each component in the composition may be 100 parts by weight. Unless otherwise specified, the percentages (including weight percentages) in this invention are based on the total weight of the composition, and "wt%" in this document refers to mass percentage.

[0044] In this document, unless otherwise stated, the reaction steps may be performed in the order described herein or not. For example, other steps may be included between reaction steps, and the order of reaction steps may be appropriately interchanged. This is something that those skilled in the art can determine based on conventional knowledge and experience. Preferably, the reaction methods described herein are performed sequentially.

[0045] In this application, when numerical intervals (i.e., numerical ranges) are involved, unless otherwise specified, the distribution of selectable numerical values ​​within the numerical interval is considered continuous, and includes the two endpoints of the numerical interval (i.e., the minimum and maximum values), as well as every numerical value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that numerical interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints, which is equivalent to directly listing every integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be merged. In other words, unless otherwise specified, the numerical ranges disclosed in this application should be understood to include any and all subranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, percentage, ratio, etc. The term "numerical interval" can be broadly included to include percentage intervals, ratio intervals, proportion intervals, etc.

[0046] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of this invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0047] This application provides a single / dual column alternating cyclic chromatography system for separation and purification to solve at least one of the following problems: (1) In traditional single-column chromatography separation, the number of qualified samples after one purification is small, requiring secondary concentration or dilution of the sample and impurities before multiple purifications, resulting in complex operation. (2) Existing simulated moving bed (SMB) systems require multiple chromatographic columns and more complex piping and valve systems to realize the function of a simulated moving bed, resulting in complex equipment structure, large footprint, high investment cost, and difficulties in cleaning and maintenance. The single / dual column alternating cyclic chromatography system for separation and purification will be described below with reference to the accompanying drawings.

[0048] The single / dual column alternating cyclic chromatography system 10 for separation and purification provided in this application embodiment is exemplarily illustrated in Figure 1, which is a schematic structural diagram of the single / dual column alternating cyclic chromatography system 10 for separation and purification provided in this application embodiment. The single / dual column alternating cyclic chromatography system 10 for separation and purification provided in this application can be used for various purposes.

[0049] To more clearly illustrate the structure of the single-column alternating cyclic chromatography system 10 used for separation and purification, the following description of the single-column alternating cyclic chromatography system 10 for separation and purification will be provided in conjunction with the accompanying drawings.

[0050] For example, referring to Figure 1, a single / dual column alternating cyclic chromatography system 10 for separation and purification includes a first column 110, a second column 120, a first detector 210, a second detector 220, a first control valve 310, a second control valve 320, a first multi-port valve 330, a second multi-port valve 340, a third multi-port valve 350, a fourth multi-port valve 360, a fifth multi-port valve 370, and a sixth multi-port valve 380. The first control valve 310, the first multi-port valve 330, the first column 110, the first detector 210, and the third multi-port valve 350 are connected sequentially. The second control valve 320, the second multi-port valve 340, the second column 120, the second detector 220, and the fourth multi-port valve 360 ​​are connected sequentially. The first multi-port valve 330 is also connected to the second control valve 320. The second multi-port valve 340 is also connected to the first control valve 310. The second multi-way valve 340 is simultaneously connected to the third multi-way valve 350, the fifth multi-way valve 370, and the sixth multi-way valve 380. The fifth multi-way valve 370 and the sixth multi-way valve 380 are also simultaneously connected to the first multi-way valve 330. The fourth multi-way valve 360 ​​is also simultaneously connected to the first multi-way valve 330, the fifth multi-way valve 370, and the sixth multi-way valve 380.

[0051] The aforementioned single / dual-column alternating cyclic chromatography system 10 for separation and purification is compact, simple in structure, requires fewer chromatographic columns, is easy to operate, and produces high purity and high yield of the separated products, enabling automated and continuous separation and purification. In the aforementioned single / dual-column alternating cyclic chromatography system 10 for separation and purification, the first chromatographic column 110 and the second chromatographic column 120 can operate independently.

[0052] In some embodiments, the first control valve 310, the second control valve 320, the first multi-way valve 330, the second multi-way valve 340, the third multi-way valve 350, the fourth multi-way valve 360, the fifth multi-way valve 370, and the sixth multi-way valve 380 are each independently selected from four-way valves.

[0053] In some embodiments, the first control valve 310 and the second control valve 320 may be selected as solvent pumps.

[0054] In some embodiments, the single / dual column alternating cyclic chromatography system 10 for separation and purification further includes a first drive pump 410. One port of the first multi-way valve 330 is connected in parallel with one port of the second multi-way valve 340 and then connected to the first control valve 310 via the first drive pump 410.

[0055] In some embodiments, the single / dual column alternating cyclic chromatography system 10 for separation and purification further includes a second drive pump 420. One port of the first multi-way valve 330 is connected in parallel with one port of the second multi-way valve 340 and then connected to the second control valve 320 via the second drive pump 420.

[0056] In some embodiments, the single / dual column alternating cyclic chromatography system 10 for separation and purification further includes a third drive pump 430. One port of the fourth multi-way valve 360, one port of the fifth multi-way valve 370, and one port of the sixth multi-way valve 380 are connected in parallel to the first multi-way valve 330 via the third drive pump 430.

[0057] In some embodiments, the single / dual column alternating cyclic chromatography system 10 for separation and purification also includes a fourth drive pump 440. The fourth drive pump 440 is connected to one of the ports of the fifth multi-port valve 370 for pumping in diluent.

[0058] In some embodiments, the single / dual column alternating cyclic chromatography system 10 for separation and purification also includes a fifth drive pump 450. The fifth drive pump 450 is connected to one of the ports of a sixth multi-port valve 380 for pumping out overflow solvent.

[0059] In some embodiments, the first detector 210 includes one or more of an ultraviolet detector, an evaporative light detector, and a differential refractive index detector.

[0060] In some embodiments, the second detector 220 includes one or more of an ultraviolet detector, an evaporative light detector, and a differential refractive index detector.

[0061] In some embodiments, referring to FIG1, the first multi-way valve 330 includes at least a first valve port connected to the first control valve 310, a second valve port connected to the second control valve 320, a third valve port connected to the first chromatographic column 110, and a fourth valve port connected to the third drive pump 430. Referring to FIG1, the numbers 1, 2, 3, and 4, arranged clockwise, represent the first, second, third, and fourth valve ports of the first multi-way valve 330, respectively.

[0062] Referring to Figure 1, the second multi-port valve 340 includes at least a first valve port connected to the first control valve 310, a second valve port connected to the second control valve 320, a third valve port connected to the second chromatographic column 120, and a fourth valve port simultaneously connected to the fifth multi-port valve 370 and the sixth multi-port valve 380. Referring to Figure 1, the numbers 1, 2, 3, and 4, arranged clockwise, represent the first, second, third, and fourth valve ports of the first multi-port valve 330, respectively. The first, second, third, and fourth valve ports of other multi-port valves are also distributed according to the above rule, and the corresponding numbers are not shown individually in Figure 1.

[0063] Referring to Figure 1, the third multi-way valve 350 includes at least a first valve port connected to the first detector 210 and used to discharge pre-impurities, a second valve port connected to the second multi-way valve 340 and the fifth multi-way valve 370, a third valve port used to discharge post-impurities, and a fourth valve port used to discharge separated samples.

[0064] Referring to Figure 1, the fourth multi-way valve 360 ​​includes at least a first valve port connected to the second detector 220 and used for discharging impurities, a second valve port used for discharging impurities, a third valve port used for discharging separated samples, and a second valve port connected simultaneously to the third drive pump 430, the fifth multi-way valve 370, and the sixth multi-way valve 380.

[0065] Referring to Figure 1, the fifth multi-way valve 370 includes at least a first valve port, a second valve port connected to the second multi-way valve 340, a third valve port for pumping in diluent, and a fourth valve port connected to both the first multi-way valve 330 and the fourth multi-way valve 360.

[0066] Referring to Figure 1, the sixth multi-way valve 380 includes at least a first valve port, a second valve port connected to the second multi-way valve 340, a third valve port for discharging overflow solvent, and a fourth valve port connected to both the first multi-way valve 330 and the fourth multi-way valve 360.

[0067] The single / dual column alternating cyclic chromatography system 10 used in this application for separation and purification enables the independent operation of the first chromatographic column 110 and the second chromatographic column 120. For example, the first control valve 310, the first drive pump 410, the first multi-way valve 330, the first chromatographic column 110, the third multi-way valve 350, and the fifth multi-way valve 370 can be sequentially connected to achieve independent single-column operation of the first chromatographic column 110. Alternatively, the second control valve 320, the second drive pump 420, the second multi-way valve 340, the second chromatographic column 120, the fourth multi-way valve 360, and the sixth multi-way valve 380 can be sequentially connected to achieve independent single-column operation of the second chromatographic column 120.

[0068] One embodiment of this application also provides a single-column and dual-column alternating cyclic chromatography method for separation and purification.

[0069] A single / dual column alternating cyclic chromatography method for separation and purification, using the aforementioned single / dual column alternating cyclic chromatography system 10 for separation and purification, includes the following method:

[0070] In the first cycle, the first drive pump 410 drives the multi-component mixture (G) through the first chromatographic column 110 for saturated loading. The second drive pump 420 elutes the first chromatographic column 110. The multi-component mixture (G) is developed in the first chromatographic column 110, sequentially separating into a pre-impurity (a), a pre-impurity and sample mixture (b), a small amount of qualified sample (c), a post-impurity and sample mixture (d), and a post-impurity section (e). Under the elution of the mobile phase, the pre-impurity (a) is discharged first through the third multi-port valve 350. The pre-impurity and sample mixture (b) is transferred to the second chromatographic column 120 by the third drive pump 430, and the small amount of qualified sample (c) is also transferred to the second chromatographic column 120 by the third drive pump 430. The post-impurity and sample mixture (d) is further separated into sections. The sample is transferred to the second column 120 via the third drive pump 430. At this time, the mixed section of the impurities and sample (b), a small amount of qualified sample, and the mixed section of the impurities and sample (d) together constitute the inner circulation sample (f). During the transfer of the inner circulation sample (f), the entire system is controlled in a closed loop state, and no additional elution solvent is required. At this stage, the fourth drive pump 440 is turned on to pump in the diluent for online dilution according to the application scenario of the sample, and the fifth drive pump 450 is turned on under the same conditions for transfer. After the transfer is completed, the first control valve 310 is switched to regenerator, and the first drive pump 410 is controlled to discharge the impurities (e) from the first column 110. Finally, the first control valve 310 is switched to elution solvent, and the first drive pump 410 is controlled to balance the first column 110, and the first cycle ends.

[0071] The second cycle begins by transferring the internal circulation sample (f) from the second column 120 into the first column 110 through the inlet for receiving the internal circulation sample (f). During the transfer of the internal circulation sample (f), the entire system is kept in a closed loop, without the need for additional elution solvent. At this point, depending on the sample application scenario, the fourth drive pump 440 is activated to pump in the diluent for online dilution, and the fifth drive pump 450 at the same flow rate is activated for transfer. After the transfer is completed, the first control valve 310 is switched to regenerator, and the first drive pump 410 is activated to discharge the impurities (e) from the second column 120. Finally, the first control valve 310 is switched to elution solvent, and the first drive pump 410 is activated to equilibrate the second column 120, thus ending the second cycle.

[0072] And repeat the first and second cycles until the inner circulation sample (f) meets the quality requirements.

[0073] In some embodiments, the impurities (a) or impurities (e) at the effluent of the first chromatographic column 110 and the second chromatographic column 120 are collected and stored respectively.

[0074] In some embodiments, the mobile phase used by the first drive pump 410 includes, but is not limited to, eluent, regeneration solution, or equilibration solution.

[0075] In some embodiments, after repeating the first and second cycles at least once, the first chromatographic column 110 and / or the second chromatographic column 120 are regenerated or equilibrated by pumping regeneration solution and equilibration solution into the first drive pump 410.

[0076] The parameters for removing precursor impurities include: the first chromatographic column 110 and the second chromatographic column 120 are in an independent state. In this invention, the flow rate of the solvent pump and the flow rate of the developing reagent are the same. In this invention, the elution time of the precursor impurities can be adjusted according to the actual application scenario. In this invention, if the precursor impurities are valuable in the actual application scenario, they can be collected and stored separately; if they are not valuable, they can be directly discharged into the waste liquid for treatment.

[0077] The parameters for the transfer of the impurity (a) and sample mixture (b) include: the first column 110 and the second column 120 are connected in a closed loop in series. The transfer time of the impurity (a) and sample mixture (b) can be adjusted according to the actual application scenario.

[0078] The parameters for transferring a small number of qualified samples include: the first chromatographic column 110 and the second chromatographic column 120 are connected in a closed loop in series. In this invention, the flow rate of the first driving pump 410 or the driving pump is consistent with the flow rate of the developing reagent. In this invention, the collection time of qualified samples can be adjusted according to the actual application scenario.

[0079] The parameters for transferring the impurity (e) to the sample mixing section (b) include: the first column 110 and the second column 120 are connected in a closed loop in series. The transfer time of the impurity (e) to the sample mixing section (b) can be adjusted according to the actual application scenario.

[0080] In this invention, the regeneration (i.e., removal of impurities) parameters include: the first chromatographic column 110 and the second chromatographic column 120 are in an independent state. In this invention, the regenerating solution enters the chromatographic column via a first drive pump 410 or a drive-driven pump. In this invention, the flow rate of the first drive pump 410 or the second drive pump 420 is the same as the flow rate of the developing reagent. In this invention, the elution intensity of the regenerating solution is greater than that of the eluent. In this invention, the regeneration time can be adjusted according to the actual application scenario.

[0081] In this invention, the equilibration parameters include: the first chromatographic column 110 and the second chromatographic column 120 are in an independent state. In this invention, the equilibration solution enters the chromatographic column via a first drive pump 410 or a second drive pump 420. In this invention, the flow rate of the first drive pump 410 or the second drive pump 420 is the same as the flow rate of the developing reagent. In this invention, the elution strength of the solution in the first drive pump 410 or the second drive pump 420 is the same as that of the developing reagent. The equilibration time can be adjusted according to the actual application scenario.

[0082] In this invention, the parameters for internal circulation sample transfer include: a first chromatographic column 110 and a second chromatographic column 120 are connected in a closed loop in series. The outlet of the second chromatographic column 120 is connected to the inlet of the first chromatographic column 110, and the outlet of the first chromatographic column 110 is connected to the inlet of the second chromatographic column 120. A circulation pump provides power to the mobile phase in the closed loop. During sample transfer, a fourth drive pump 440 is activated for online dilution, and a fifth drive pump 450 is activated for online overflow, depending on the application scenario. The flow rates of the circulation pump, the fourth drive pump 440, and the fifth drive pump 450 can be adjusted according to the actual application scenario. The conditions of the fourth drive pump 440 and the fifth drive pump 450 should be kept consistent.

[0083] In this invention, to achieve an efficient and continuous separation and purification process for multi-component mixtures, the first and second cycles can be performed at least twice. The first and second cycles are referred to as a large cycle, and subsequent steps involve repeating the large cycle multiple times until the inner cycle sample meets the quality requirements. Subsequent sample processing steps are the same as those for the first and second cycles, and so on.

[0084] Example 1

[0085] In this embodiment, oil extracted from animals is used as raw material. The oil, with a purity of 80%, is dissolved in an aqueous methanol solution to prepare a sample solution with a concentration of 100 mg / mL. Figure 2 shows the liquid chromatography chromatogram of the prepared oil. In Figure 2, the horizontal axis represents time (in minutes), and the vertical axis represents the absorbance value (in mAu).

[0086] The sample solution was separated and purified using the single and dual column alternating cyclic chromatography system 10 described above.

[0087] In the single-column alternating cyclic chromatography system 10 used for separation and purification, the dimensions of the first chromatographic column 110 are both 10 mm in diameter and 250 mm in length. The packing material of the first chromatographic column 110 is C18 packing material with a particle size of 30 μm (C18 packing material is sourced from Jiangsu Hanbang Technology Co., Ltd.).

[0088] The single / dual column alternating cyclic chromatography method for separation and purification includes the following steps:

[0089] In the first cycle, the first drive pump 410 drives the multi-component mixture (G) through the first chromatographic column 110 for saturated loading. The second drive pump 420 elutes the first chromatographic column 110. The multi-component mixture (G) is developed in the first chromatographic column 110, sequentially separating into a pre-impurity (a), a pre-impurity and sample mixture (b), a small amount of qualified sample (c), a post-impurity and sample mixture (d), and a post-impurity section (e). Under the elution of the mobile phase, the pre-impurity (a) is discharged first through the third multi-port valve 350. The pre-impurity and sample mixture (b) is transferred to the second chromatographic column 120 by the third drive pump 430, and the small amount of qualified sample (c) is also transferred to the second chromatographic column 120 by the third drive pump 430. The post-impurity and sample mixture (d) is further separated into sections. The sample is transferred to the second column 120 via the third drive pump 430. At this time, the mixed section of the impurities and sample (b), a small amount of qualified sample, and the mixed section of the impurities and sample (d) together constitute the inner circulation sample (f). During the transfer of the inner circulation sample (f), the entire system is controlled in a closed loop state, and no additional elution solvent is required. At this stage, the fourth drive pump 440 is turned on to pump in the diluent for online dilution according to the application scenario of the sample, and the fifth drive pump 450 is turned on under the same conditions for transfer. After the transfer is completed, the first control valve 310 is switched to regenerator, and the first drive pump 410 is controlled to discharge the impurities (e) from the first column 110. Finally, the first control valve 310 is switched to elution solvent, and the first drive pump 410 is controlled to balance the first column 110, and the first cycle ends.

[0090] The second cycle begins by transferring the internal circulation sample (f) from the second column 120 into the first column 110 through the inlet for receiving the internal circulation sample (f). During the transfer of the internal circulation sample (f), the entire system is kept in a closed loop, without the need for additional elution solvent. At this point, depending on the sample application scenario, the fourth drive pump 440 is activated to pump in the diluent for online dilution, and the fifth drive pump 450 at the same flow rate is activated for transfer. After the transfer is completed, the first control valve 310 is switched to regenerator, and the first drive pump 410 is activated to discharge the impurities (e) from the second column 120. Finally, the first control valve 310 is switched to elution solvent, and the first drive pump 410 is activated to equilibrate the second column 120, thus ending the second cycle.

[0091] And repeat the first and second cycles until the inner circulation sample (f) meets the quality requirements.

[0092] In the method of this embodiment, the loading parameters include: the first chromatographic column 110 and the second chromatographic column 120 are in an independent state, the flow rate of the loading pump is 3 mL / min, and the loading time is 1 min.

[0093] The parameters for the developing layer include: the first column 110 and the second column 120 are independent; the developing reagent is 96% ethanol (volume fraction); the solvent pump flow rate for the developing reagent is 5 mL / min; and the developing time is 5 min.

[0094] The parameters for the transfer of the pre-impurities and sample mixture, the parameters for the transfer of a small amount of qualified sample, and the parameters for the transfer of the post-impurities and sample mixture all include: the first column 110 and the second column 120 are connected in a closed loop in series, the eluent is the solvent in the second column 120, and the circulation pump flow rate is 5 mL / min; the parameters for the transfer of the internal circulating sample in the first column 110 to the fourth drive pump 440 and the fifth drive pump 450 in the second column 120 include: the flow rate of the fourth drive pump 440 and the fifth drive pump 450 is 0.2 mL / min, and the diluent is purified water.

[0095] The regeneration and equilibration parameters include: the first column 110 and the second column 120 are in an independent state; the equilibration solution and developing reagent are the same; the regeneration solution is pure ethanol; the flow rate of the first drive pump 410 for the eluent is 5 mL / min; the regeneration and equilibration times are 3 min each; the first cycle ends.

[0096] At the start of the second cycle, the internal circulation sample of the second column 120 is transferred to the first column 110 through the inlet for receiving the internal circulation sample. During the transfer of the internal circulation sample, the entire system is in a closed loop state, and no additional elution solvent is required. At this time, the fourth drive pump 440 is started for online dilution according to the sample application scenario, and the fifth drive pump 450 at the same flow rate is started for transfer. After the transfer is completed, the solvent selection valve V01 is switched to regenerator and the P01 pump is started to discharge the impurities from the second column 120. Finally, the solvent selection valve V01 is switched to elution solvent and the P01 pump is started to equilibrate the second column 120, and the second cycle ends.

[0097] The first and second cycles are called a large cycle. Subsequent steps involve repeating the large cycle multiple times until the inner cycle sample meets the quality requirements. All subsequent steps are the same as the large cycle steps described above, and so on.

[0098] The parameters for internal circulation sample transfer include: the first column 110 and the second column 120 are connected in a closed loop in series; the eluent is the solvent in the first column 110; and the circulation pump flow rate is 5 mL / min. The parameters for internal circulation sample transfer from the second column 120 to the fourth drive pump 440 and the fifth drive pump 450 in the first column 110 include: the flow rate of the fourth drive pump 440 and the fifth drive pump 450 is 0.2 mL / min; and the diluent is purified water.

[0099] The regeneration and equilibration parameters include: the first column 110 and the second column 120 are in an independent state; the equilibration solution and developing reagent are the same; the regeneration solution is pure ethanol; the flow rate of the first drive pump 410 for the eluent is 5 mL / min; and the regeneration and equilibration times are 3 min each.

[0100] After four large cycles, a qualified sample was collected from the second valve port of the second column 120.

[0101] The purity of the collected qualified samples was determined by gas chromatography. The gas chromatography conditions were as follows: instrument model: Agilent GC8860; column: Agilent XB-WAX 25m×320um×0.15μm; injection volume: 1μL; detection conditions: injection port 280℃; split ratio 25:1; split flow rate 30mL / min; column temperature: initial 150℃, rate ramping to 270℃ at 2.5℃ / min, hold time 5min; FID detector: 270℃.

[0102] The obtained gas chromatogram is shown in Figure 3. Figure 3 is a detection spectrum of the oil after separation and purification using a single / dual column alternating cyclic chromatography system 10 according to an embodiment of the present invention. As shown in Figure 3, the detection spectrum of the qualified sample collected after preparation shows that the raw material undergoes multiple cyclic transfers, finally achieving good separation between the sample and impurities. The purity of the qualified product can be greater than 99%, and the calculated yield is greater than 90%.

[0103] In summary, this invention achieves continuous and automated cyclic operation of dual-column chromatography by reducing the number of chromatographic columns and utilizing the countercurrent operation of a simulated moving bed (SMB). Its main advantages lie in using two chromatographic columns to achieve the functions of simulated moving bed countercurrent operation and continuous operation. It can truly and continuously separate and purify multi-component mixtures with lower investment costs. It achieves advantages such as simpler equipment structure, flexible and convenient operation, higher separation efficiency and yield, automated continuous operation, saving time and labor costs, low investment cost, easy maintenance and cleaning, and significantly reducing the amount of eluent used.

[0104] In the above embodiments, the descriptions of each embodiment have different focuses. For parts not described in detail in a certain embodiment, please refer to the relevant descriptions in other embodiments.

[0105] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0106] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these modifications and improvements all fall within the scope of protection of the present invention. Therefore, the scope of protection of this patent should be determined by the appended claims.

Claims

1. A single / dual column alternating cyclic chromatography system for separation and purification, characterized in that, The system includes a first chromatographic column, a second chromatographic column, a first detector, a second detector, a first control valve, a second control valve, a first multi-port valve, a second multi-port valve, a third multi-port valve, a fourth multi-port valve, a fifth multi-port valve, and a sixth multi-port valve. The first control valve, the first multi-port valve, the first chromatographic column, the first detector, and the third multi-port valve are connected sequentially. The second control valve, the second multi-port valve, the second chromatographic column, the second detector, and the fourth multi-port valve are also connected sequentially. The first multi-port valve is further connected to the second control valve, and the second multi-port valve is also connected to the first control valve. The second multi-port valve is simultaneously connected to the third, fifth, and sixth multi-port valves. The fifth and sixth multi-port valves are also simultaneously connected to the first multi-port valve, and the fourth multi-port valve is also simultaneously connected to the first, fifth, and sixth multi-port valves.

2. The single / dual column alternating cyclic chromatography system for separation and purification according to claim 1, characterized in that, The first control valve, the second control valve, the first multi-way valve, the second multi-way valve, the third multi-way valve, the fourth multi-way valve, the fifth multi-way valve, and the sixth multi-way valve are each independently selected from four-way valves.

3. The single / dual column alternating cyclic chromatography system for separation and purification according to claim 1, characterized in that, The single / dual column alternating cyclic chromatography system for separation and purification further includes a first drive pump, wherein one of the valve ports of the first multi-way valve and one of the valve ports of the second multi-way valve are connected in parallel and then connected to the first control valve via the first drive pump.

4. The single / dual column alternating cyclic chromatography system for separation and purification according to claim 1, characterized in that, The single / dual column alternating cyclic chromatography system for separation and purification further includes a second drive pump. One of the valve ports of the first multi-way valve and one of the valve ports of the second multi-way valve are connected in parallel and then connected to the second control valve through the second drive pump.

5. The single / dual column alternating cyclic chromatography system for separation and purification according to claim 1, characterized in that, The single / dual column alternating cyclic chromatography system for separation and purification further includes a third drive pump. One of the valve ports of the fourth multi-port valve, one of the valve ports of the fifth multi-port valve, and one of the valve ports of the sixth multi-port valve are connected in parallel and then connected to the first multi-port valve through the third drive pump.

6. The single / dual column alternating cyclic chromatography system for separation and purification according to claim 1, characterized in that, The single / dual column alternating cyclic chromatography system for separation and purification also includes a fourth drive pump, which is connected to one of the ports of the fifth multi-port valve for pumping in diluent. And / or, the single / dual column alternating cyclic chromatography system for separation and purification further includes a fifth drive pump connected to one of the ports of the sixth multi-port valve for pumping out overflow solvent.

7. The single / dual column alternating cyclic chromatography system for separation and purification according to any one of claims 1 to 6, characterized in that, The first detector includes one or more of the following: an ultraviolet detector, an evaporative light detector, and a differential refractive index detector; And / or, the second detector includes one or more of the following: an ultraviolet detector, an evaporative light detector, and a differential refractive index detector.

8. The single / dual column alternating cyclic chromatography system for separation and purification according to claim 1, characterized in that, The first multi-port valve includes at least a first valve port connected to the first control valve, a second valve port connected to the second control valve, a third valve port connected to the first chromatographic column, and a fourth valve port connected to the third drive pump. The second multi-way valve includes at least a first valve port connected to the first control valve, a second valve port connected to the second control valve, a third valve port connected to the second chromatographic column, and a fourth valve port simultaneously connected to the fifth multi-way valve and the sixth multi-way valve; The third multi-way valve includes at least a first valve port connected to the first detector and used to discharge pre-impurities, a second valve port connected to both the second and fifth multi-way valves, a third valve port used to discharge post-impurities, and a fourth valve port used to discharge separated samples. The fourth multi-way valve includes at least a first valve port connected to the second detector and used for discharging impurities, a second valve port used for discharging impurities, a third valve port used for discharging separated samples, and a second valve port connected simultaneously to the third drive pump, the fifth multi-way valve, and the sixth multi-way valve. The fifth multi-way valve includes at least a first valve port, a second valve port connected to the second multi-way valve, a third valve port for pumping in diluent, and a fourth valve port connected to both the first and fourth multi-way valves. The sixth multi-way valve includes at least a first valve port, a second valve port connected to the second multi-way valve, a third valve port for discharging overflow solvent, and a fourth valve port connected to both the first and fourth multi-way valves.

9. A single / dual column alternating cyclic chromatography method for separation and purification, characterized in that, Using the single / dual column alternating cyclic chromatography system for separation and purification according to any one of claims 1 to 8, the method comprises the following steps: In the first cycle, the multi-component mixture (G) is saturated and loaded onto the first chromatographic column. The column is then eluted, and the multi-component mixture (G) is developed and sequentially separated into a pre-impurity (a), a pre-impurity and sample mixture (b), a small amount of acceptable sample (c), a post-impurity and sample mixture (d), and a post-impurity section (e). Under elution with the mobile phase, the pre-impurity (a) is discharged first through the third multi-port valve. The pre-impurity and sample mixture (b) is transferred to the second chromatographic column, and the small amount of acceptable sample (c) is transferred to... In the second column, the impurity-sample mixture (d) is transferred to the second column. The impurity-sample mixture (b), a small amount of qualified sample, and the impurity-sample mixture (d) together form the internal circulation sample (f). During the transfer of the internal circulation sample (f), the entire system is controlled in a closed loop state, without the need to add additional elution solvent. At this stage, diluent is pumped in for online dilution according to the application scenario of the sample. The first control valve switches to regenerator to discharge the impurity (e) from the first column. The first control valve switches to elution solvent to equilibrate the first column, and the first cycle ends. The second cycle begins by transferring the internal circulation sample (f) from the second column to the first column through the inlet of the first column used to receive the internal circulation sample (f). During the transfer of the internal circulation sample (f), the entire system is kept in a closed loop state without the need for additional elution solvent. At this time, diluent is pumped in for online dilution according to the sample application scenario. The first control valve is switched to regenerant, and the first drive pump is turned on to discharge the impurities (e) from the second column. The first control valve is switched to elution solvent, and the first drive pump is turned on to equilibrate the second column. The second cycle ends. And repeat the first and second cycles until the inner circulation sample (f) meets the quality requirements.

10. The single / dual column alternating cyclic chromatography method for separation and purification according to claim 9, characterized in that, The single / dual column alternating cyclic chromatography method for separation and purification also satisfies at least one of the following conditions: (1) Collect and store the impurities (a) or impurities (e) at the effluent of the first chromatographic column and the second chromatographic column respectively; (2) The mobile phase includes, but is not limited to, elution buffer, regeneration buffer or equilibration buffer; (3) After repeating the first and second cycles at least once, the first chromatographic column and / or the second chromatographic column are regenerated or balanced by pumping regeneration solution and equilibration solution into the first drive pump.

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