Single-column and double-column alternate circulation chromatographic system for separation and purification and method thereof

Abstract

The invention discloses a single-column and double-column alternate circulation chromatographic system for separation and purification and a method of the single-column and double-column alternate circulation chromatographic system. 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, and the first control valve, the first multi-way valve, the first chromatographic column, the first detector and the third multi-way valve are sequentially connected. The second control valve, the second multi-way valve, the second chromatographic column, the second detector and the fourth multi-way valve are sequentially connected, the first multi-way valve is connected to the second control valve, the second multi-way valve is further connected to the first control valve, and the second multi-way valve is connected with 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 further connected with the first multi-way valve, and the fourth multi-way valve is further connected with the first multi-way valve, the fifth multi-way valve and the sixth multi-way valve. The equipment is simple in structure and high in separation efficiency and yield.

Description

Technical Field

[0001] The present application relates to the technical field of chromatography detection, and in particular to a single-column and double-column alternating circulation chromatography system and method for separation and purification. Background Art

[0002] In the field of chromatography detection, the traditional single-column chromatography process is usually operated intermittently, requiring continuous feeding, elution, collection and other steps. The operation is cumbersome, time-consuming, and requires a large amount of eluent, resulting in high production costs. In addition, in the traditional single-column chromatography separation process, there are few qualified samples after one purification, and the sample and impurity cross-section needs to be concentrated or diluted for multiple purifications. The existing simulated moving bed SMB (Simulated Moving Bed) realizes the countercurrent between the stationary phase and the mobile phase through its special process equipment and automatic control technology, so that the contact between the separated substances in the system and the stationary phase is more sufficient, so that different components in the mixture can be effectively separated, and higher purity samples can be obtained with less eluent consumption. The system can also be operated automatically and continuously, reducing labor costs. However, the traditional simulated moving bed SMB usually requires multiple chromatographic columns and more complex piping and valve systems to realize the function of the simulated moving bed. The equipment has a complex structure, large floor space, high investment cost, and is difficult to clean and has high maintenance costs. Summary of the invention

[0003] Based on this, it is necessary to provide a single-column and double-column alternating circulation chromatography system for separation and purification. The single-column and double-column alternating circulation chromatography system for separation and purification of the present invention has compact equipment, simple structure, small number of chromatographic columns, easy operation, high purity and high yield of separated products, and can realize automated and continuous separation and purification.

[0004] An embodiment of the present application provides a single-column and double-column alternating circulation chromatography system for separation and purification.

[0005] A single-column and double-column alternating circulation chromatography system for separation and purification, comprising 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, wherein 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 also 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 simultaneously 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 simultaneously connected to the first multi-way valve, and the fourth multi-way valve is also simultaneously connected to the first multi-way valve, the fifth multi-way valve and the sixth multi-way valve.

[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 independently selected from four-way valves.

[0007] In some embodiments, the single-column and double-column alternating circulation chromatography system for separation and purification also includes a first driving pump, and one of the valve ports of the first multi-way valve is connected in parallel with one of the valve ports of the second multi-way valve and then connected to the first control valve through the first driving pump.

[0008] In some embodiments, the single-column and double-column alternating circulation chromatography system for separation and purification also includes a second driving pump, and one of the valve ports of the first multi-way valve is connected in parallel with one of the valve ports of the second multi-way valve and then connected to the second control valve through the second driving pump.

[0009] In some embodiments, the single-column and double-column alternating circulation chromatography system for separation and purification also includes a third driving pump, and one of the valve ports of the fourth multi-way valve, one of the valve ports of the fifth multi-way valve, and one of the valve ports of the sixth multi-way valve are connected in parallel and connected to the first multi-way valve through the third driving pump.

[0010] In some embodiments, the single-column and double-column alternating circulation chromatography system for separation and purification further comprises a fourth driving pump, which is connected to one of the valve ports of the fifth multi-way valve for pumping in a diluent.

[0011] In some embodiments, the single-column and double-column alternating circulation chromatography system for separation and purification further comprises a fifth driving pump, which is connected to one of the valve ports of the sixth multi-way 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-way valve at least includes 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;

[0015] The second multi-way valve at least comprises 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 connected to both the fifth multi-way valve and the sixth multi-way valve;

[0016] The third multi-way valve at least includes a first valve port connected to the first detector and used to discharge the front impurities, a second valve port connected to the second multi-way valve and the fifth multi-way valve, a third valve port used to discharge the rear impurities, and a fourth valve port used to discharge the separated sample;

[0017] The fourth multi-way valve at least includes a first valve port connected to the second detector and used to discharge the residual impurities, a second valve port used to discharge the residual impurities, a third valve port used to discharge the separated sample, and a second valve port connected to the third driving pump, the fifth multi-way valve and the sixth multi-way valve at the same time;

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

[0019] The sixth multi-way valve at least includes 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 multi-way valve and the fourth multi-way valve.

[0020] An embodiment of the present application also provides a single-column and double-column alternating cycle chromatography method for separation and purification.

[0021] A single-column and double-column alternating circulation chromatography method for separation and purification, using the above-mentioned single-column and double-column alternating circulation chromatography system for separation and purification, comprises the following steps:

[0022] In a first cycle, the multi-component mixture (G) is saturatedly loaded through the first chromatographic column, and the first chromatographic column is eluted. The multi-component mixture (G) is layered in the first chromatographic column and sequentially divided into a pre-impurity (a), a pre-impurity and sample mixed section (b), a small amount of qualified sample section (c), a post-impurity and sample mixed section (d), and a post-impurity section (e); under the elution of the first chromatographic column with the mobile phase, the pre-impurity (a) is first discharged by the third multi-way valve; the pre-impurity and sample mixed section (b) is transferred to the second chromatographic column, and the small amount of qualified sample section (c) is transferred to the The second chromatographic column; the post-impurity and sample mixing section (d) is transferred to the second chromatographic column, and the pre-impurity and sample mixing section (b), a small amount of qualified sample section and the post-impurity and sample mixing section (d) together constitute the inner circulation sample (f). During the transfer of the inner circulation sample (f), the entire system is controlled to be in a closed loop state, and no additional elution solvent is required. At this stage, a diluent is pumped in for online dilution according to the application scenario of the sample; the first control valve is switched to a regenerant to discharge the post-impurity (e) of the first chromatographic column; the first control valve is switched to an elution solvent to balance the first chromatographic column, and the first cycle ends;

[0023] The second cycle begins, the inner circulation sample (f) of the second chromatographic column is transferred to the first chromatographic column through the inlet of the first chromatographic column for receiving the inner circulation sample (f), and the whole system is controlled to be in a closed loop state during the transfer of the inner circulation sample (f), and no additional elution solvent is needed. At this time, a diluent is pumped in for online dilution according to the sample application scenario; the first control valve is switched to a regeneration agent, and the first drive pump is turned on to discharge the impurities (e) after the second chromatographic column; the first control valve is switched to an elution solvent, and the first drive pump is turned on to balance the second chromatographic column, and the second cycle ends;

[0024] And, repeating the first cycle and the second cycle until the inner cycle sample (f) meets the quality requirements.

[0025] In some embodiments, the front impurities (a) or the rear impurities (e) at the outer discharge 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 driving pump includes but is not limited to an eluent, a regeneration solution, or an equilibrium solution.

[0027] In some embodiments, after repeating the first cycle and the second cycle at least once, the first chromatographic column and / or the second chromatographic column is regenerated or balanced by pumping in a regeneration liquid and a balancing liquid through a first driving pump.

[0028] The single-column and double-column alternating circulation chromatography system for separation and purification is compact, simple in structure, requires a small number of chromatography columns, is easy to operate, and has high purity and yield of the separated products, and can realize automated and continuous separation and purification.

[0029] The present invention reduces the number of chromatographic columns and utilizes the countercurrent operation of a simulated moving bed (SMB) to achieve continuous and automated cyclic operation of a double-column chromatography. The main advantage of the present invention is that two chromatographic columns are used to achieve the countercurrent operation and continuous operation functions of a simulated moving bed, and a multi-component mixture can be truly and continuously separated and purified at a relatively low investment cost, thereby achieving the advantages of simpler equipment structure, flexible and convenient operation, higher separation efficiency and yield, automated and continuous operation, time and labor cost savings, low investment cost, easy maintenance and cleaning, and the ability to significantly reduce the amount of eluent used. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments of the present application, and those skilled in the art can obtain other drawings based on these drawings without creative work.

[0031] In order to more completely understand the present application and its beneficial effects, the following description will be given in conjunction with the accompanying drawings. In the following description, the same reference numerals represent the same parts.

[0032] FIG1 is a control schematic diagram of a single-column and double-column alternating circulation chromatography system for separation and purification according to an embodiment of the present invention;

[0033] FIG2 is a spectrum of a certain oil liquid chromatography preparation according to an embodiment of the present invention;

[0034] FIG3 is a detection spectrum of oil separation and purification by the single-column and double-column alternating circulation chromatography system for separation and purification according to an embodiment of the present invention.

[0035] Description of Reference Numerals

[0036] 10. A single-column and double-column alternating circulation chromatography system for separation and purification; 110. A first chromatography column; 120. A second chromatography column; 210. A first detector; 220. A second detector; 310. A first control valve; 320. A second control valve; 330. A first multi-way valve; 340. A second multi-way valve; 350. A third multi-way valve; 360. A fourth multi-way valve; 370. A fifth multi-way valve; 380. A sixth multi-way valve; 410. A first driving pump; 420. A second driving pump; 430. A third driving pump; 440. A fourth driving pump; 450. A fifth driving pump. DETAILED DESCRIPTION

[0037] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific embodiments of the present invention are described in detail below in conjunction with the accompanying drawings. In the following description, many specific details are set forth to facilitate a full understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below.

[0038] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as limiting the present invention.

[0039] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0040] In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may mean that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, a first feature being "above", "above" or "above" a second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being "below", "below" or "below" a second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

[0041] In the description of the present invention, "several" means more than one, "many" means more than two, "greater than", "less than", "exceed", etc. are understood to exclude the number itself, and "above", "below", "within", etc. are understood to include the number itself. If there is a description of "first" or "second", it is only used for the purpose of distinguishing the technical features, and cannot be understood as indicating or implying the relative importance or implicitly indicating the number of the indicated technical features or implicitly indicating the order of the indicated technical features.

[0042] In this article, "optionally", "optional", "optional" means optional, that is, it means to be selected from any of the two parallel solutions of "yes" or "no". If there are multiple "optional" in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent. In this application, the descriptions such as "optionally contain" and "optionally include" mean "contain or not contain".

[0043] In the present invention, unless otherwise stated, the sum of the parts of each component in the composition can be 100 parts by weight. Unless otherwise specified, the basis of the percentage (including weight percentage) of the present invention is the total weight of the composition, and "wt%" herein means mass percentage.

[0044] In this article, unless otherwise stated, each reaction step may be carried out in the order described herein or may not be carried out in the order described herein. For example, other steps may be included between each reaction step, and the order of the reaction steps may also be appropriately swapped. This can be determined by the technician based on common knowledge and experience. Preferably, the reaction method herein is carried out sequentially.

[0045] In the present application, when it comes to a numerical interval (i.e., a numerical range), unless otherwise specified, the distribution of the optional numerical values ​​in the numerical interval is considered to be continuous, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical interval, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to an integer in the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints, which is equivalent to directly listing each integer. When multiple numerical ranges are provided to describe features or characteristics, these numerical ranges can be combined. In other words, unless otherwise specified, the numerical range disclosed in the present application should be understood to include any and all sub-ranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows for a broad range of quantitative intervals such as percentage intervals, ratio intervals, and ratio intervals.

[0046] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more of the related listed items.

[0047] The embodiment of the present application provides a single-column and double-column alternating circulation chromatography system for separation and purification to solve at least one of the following problems: (1) In the traditional single-column chromatography separation process, there are few qualified samples after one purification, and the sample and the impurity cross-portion need to be concentrated or diluted for a second time and then purified multiple times, which has the problem of complex operation. (2) The existing simulated moving bed SMB (Simulated Moving Bed) requires multiple chromatographic columns and more complex piping and valve systems to realize the function of the simulated moving bed. The equipment has a complex structure, large floor space, high investment cost, and is difficult to clean and has high maintenance costs. The following will describe the single-column and double-column alternating circulation chromatography system for separation and purification in conjunction with the accompanying drawings.

[0048] The single-column and double-column alternating circulation chromatography system 10 for separation and purification provided in the embodiment of the present application is exemplarily shown in FIG1 , which is a schematic diagram of the structure of the single-column and double-column alternating circulation chromatography system 10 for separation and purification provided in the embodiment of the present application. The single-column and double-column alternating circulation chromatography system 10 for separation and purification of the present application can be used for purposes.

[0049] In order to more clearly illustrate the structure of the single-column and double-column alternating circulation chromatography system 10 for separation and purification, the single-column and double-column alternating circulation chromatography system 10 for separation and purification will be introduced below with reference to the accompanying drawings.

[0050] Exemplarily, please refer to FIG. 1, a single-column alternating circulation chromatography system 10 for separation and purification includes a first chromatographic column 110, a second chromatographic column 120, a first detector 210, a second detector 220, a first control valve 310, a second control valve 320, a first multi-way valve 330, a second multi-way valve 340, a third multi-way valve 350, a fourth multi-way valve 360, a fifth multi-way valve 370, and a sixth multi-way valve 380. Among them, the first control valve 310, the first multi-way valve 330, the first chromatographic column 110, the first detector 210, and the third multi-way valve 350 are connected in sequence. The second control valve 320, the second multi-way valve 340, the second chromatographic column 120, the second detector 220, and the fourth multi-way valve 360 ​​are connected in sequence. The first multi-way valve 330 is also connected to the second control valve 320. The second multi-way 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 single-column and double-column alternating circulation chromatography system 10 for separation and purification is compact, simple in structure, has a small number of chromatography columns, is easy to operate, and has high purity and high yield of the separated product, and can realize automated and continuous separation and purification. In the single-column and double-column alternating circulation chromatography system 10 for separation and purification, the first chromatography column 110 and the second chromatography column 120 can be operated 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 independently selected from four-way valves.

[0053] In some embodiments, the first control valve 310 and the second control valve 320 can select a solvent pump respectively.

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

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

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

[0057] In some embodiments, the single-column and double-column alternating cycle chromatography system 10 for separation and purification further includes a fourth driving pump 440. The fourth driving pump 440 is connected to one of the valve ports of the fifth multi-way valve 370 for pumping in a diluent.

[0058] In some embodiments, the single-column and double-column alternating cycle chromatography system 10 for separation and purification further includes a fifth driving pump 450. The fifth driving pump 450 is connected to one of the valve ports of the sixth multi-way valve 380 to pump out the 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, as shown in 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 driving pump 430. As shown in FIG1 , the numbers 1, 2, 3, and 4 distributed clockwise are respectively shown in the first multi-way valve 330, representing the first valve port, the second valve port, the third valve port, and the fourth valve port of the first multi-way valve 330, respectively.

[0062] As shown in FIG1 , the second multi-way valve 340 at least includes 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 connected to both the fifth multi-way valve 370 and the sixth multi-way valve 380. As shown in FIG1 , the numbers 1, 2, 3, and 4 are respectively shown in a clockwise distribution in the second multi-way valve 340, representing the first valve port, the second valve port, the third valve port, and the fourth valve port of the first multi-way valve 330, respectively. The first valve port, the second valve port, the third valve port, and the fourth valve port of other multi-way valves are also distributed according to the above rules, and the corresponding numbers are not shown one by one in FIG1 .

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

[0064] 1 , the fourth multi-way valve 360 ​​includes at least a first valve port connected to the second detector 220 and used to discharge residual impurities, a second valve port used to discharge residual impurities, a third valve port used to discharge separated samples, and a second valve port connected to the third driving pump 430, the fifth multi-way valve 370 and the sixth multi-way valve 380 at the same time.

[0065] 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] 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-column and double-column alternating circulation chromatography system 10 for separation and purification of the present application can realize 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 are connected in sequence to realize the independent single-column operation of the first chromatographic column 110. Or. 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 are connected in sequence to realize the independent single-column operation of the second chromatographic column 120.

[0068] An embodiment of the present application also provides a single-column and double-column alternating cycle chromatography method for separation and purification.

[0069] A single-column and double-column alternating circulation chromatography method for separation and purification, using the above-mentioned single-column and double-column alternating circulation chromatography system 10 for separation and purification, includes the following method:

[0070] A first cycle is performed, the first driving pump 410 is controlled to drive the multi-component mixture (G) to be saturatedly loaded through the first chromatographic column 110, and the second driving pump 420 is controlled to elute the first chromatographic column 110. The multi-component mixture (G) is layered in the first chromatographic column 110 and is sequentially divided into a front impurity (a), a front impurity and sample mixing section (b), a small amount of qualified sample section (c), a rear impurity and sample mixing section (d) and a rear impurity section (e); under the elution of the mobile phase from the first chromatographic column 110, the front impurity (a) is first discharged by the third multi-way valve 350; the front impurity and sample mixing section (b) is transferred to the second chromatographic column 120 through the third driving pump 430, and a small amount of qualified sample section (c) is transferred to the second chromatographic column 120 through the third driving pump 430; the rear impurity and sample mixing section (d ) is transferred to the second chromatographic column 120 through the third driving pump 430. At this time, the front impurity and sample mixing section (b) and a small amount of qualified sample section and the rear impurity and sample mixing section (d) together constitute the inner circulation sample (f). During the transfer of the inner circulation sample (f), the entire system is controlled to be in a closed-loop state, and no additional elution solvent is required. At this stage, according to the application scenario of the sample, the fourth driving pump 440 is turned on to pump in the diluent for online dilution and the fifth driving pump 450 is turned on under the same conditions to perform the transfer. After the transfer is completed, the first control valve 310 is switched to the regeneration agent, and the first driving pump 410 is controlled to discharge the rear impurities (e) of the first chromatographic column 110; finally, the first control valve 310 is switched to the elution solvent, and the first driving pump 410 is controlled to balance the first chromatographic column 110, and the first cycle is completed.

[0071] At the beginning of the second cycle, the inner circulation sample (f) of the second chromatographic column 120 is transferred to the first chromatographic column 110 through the inlet of the first chromatographic column 110 for receiving the inner circulation sample (f). During the transfer of the inner circulation sample (f), the entire system is controlled to be in a closed-loop state without adding additional elution solvent. At this time, according to the sample application scenario, the fourth drive pump 440 is turned on to pump in the diluent for online dilution and the fifth drive pump 450 with the same flow rate is turned on at the same time for transfer. After the transfer is completed, the first control valve 310 is switched to the regeneration agent, and the first drive pump 410 is turned on to discharge the impurities (e) after the second chromatographic column 120; finally, the first control valve 310 is switched to the elution solvent, and the first drive pump 410 is turned on to balance the second chromatographic column 120, and the second cycle ends.

[0072] And, repeating the first cycle and the second cycle until the inner cycle sample (f) meets the quality requirements.

[0073] In some embodiments, the front impurities (a) or the rear impurities (e) at the outer discharge 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 driving pump 410 includes but is not limited to an eluent, a regeneration solution, or an equilibrium solution.

[0075] In some embodiments, after repeating the first cycle and the second cycle at least once, the regeneration liquid and the balancing liquid are pumped into the first driving pump 410 to regenerate or balance the first chromatographic column 110 and / or the second chromatographic column 120 .

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

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

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

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

[0080] In the present invention, the parameters of regeneration (i.e., post-impurity removal) include: the first chromatographic column 110 and the second chromatographic column 120 are in an independent state. In the present invention, the regeneration liquid enters the chromatographic column through the first drive pump 410 or the drive drive pump. In the present invention, the flow rate of the first drive pump 410 or the second drive pump 420 is consistent with the flow rate of the spreading agent. In the present invention, the elution strength of the regeneration liquid is greater than the elution strength of the eluent. In the present invention, the regeneration time can be adjusted according to the actual application scenario.

[0081] In the present invention, the parameters of equilibrium include: the first chromatographic column 110 and the second chromatographic column 120 are in an independent state. In the present invention, the equilibrium liquid enters the chromatographic column through the first driving pump 410 or the second driving pump 420. In the present invention, the flow rate of the first driving pump 410 or the second driving pump 420 is consistent with the flow rate of the spreading agent. In the present invention, the elution strength of the first driving pump 410 or the second driving pump 420 solution is consistent with the spreading agent. The equilibrium time can be adjusted according to the actual application scenario.

[0082] In the present invention, the parameters of the inner circulation sample transfer include: the first chromatographic column 110 and the second chromatographic column 120 are connected in series in a closed loop. In the present invention, 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. In the present invention, the circulation pump provides power for the mobile phase in the closed loop connection. In the present invention, the inner circulation sample is turned on the fourth drive pump 440 for online dilution according to the application scenario of the sample during the transfer process and the fifth drive pump 450 is turned on for online overflow at the same time. In the present invention, the flow rate of the circulation pump and the fourth drive pump 440 and the fifth drive pump 450 can be adjusted according to the actual application scenario. In the present invention, the conditions of the fourth drive pump 440 and the fifth drive pump 450 should be kept consistent.

[0083] In the present invention, in order to realize the efficient and continuous separation and purification process of the multi-component mixture, the number of the first cycle and the second cycle can be ≥2 times, the first cycle and the second cycle are called a large cycle, and the subsequent steps are to repeat the large cycle multiple times until the inner cycle sample meets the quality requirements. The subsequent sample processing steps are the same as the first cycle and the second cycle steps, and so on.

[0084] Example 1

[0085] In this embodiment, oil extracted from animals is used as raw material, and a sample solution with a concentration of 100 mg / mL is prepared by dissolving the oil with a purity of 80% in methanol water solution. FIG2 is a spectrum of oil liquid chromatography preparation, in which the horizontal axis is time "min" and the vertical axis is absorption value "mAu".

[0086] The sample solution is separated and purified using the above-mentioned single-column and double-column alternating circulation chromatography system 10 for separation and purification.

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

[0088] The single-column and double-column alternating cycle chromatography method for separation and purification comprises the following steps:

[0089] A first cycle is performed, the first driving pump 410 is controlled to drive the multi-component mixture (G) to be saturatedly loaded through the first chromatographic column 110, and the second driving pump 420 is controlled to elute the first chromatographic column 110. The multi-component mixture (G) is layered in the first chromatographic column 110 and is sequentially divided into a front impurity (a), a front impurity and sample mixing section (b), a small amount of qualified sample section (c), a rear impurity and sample mixing section (d) and a rear impurity section (e); under the elution of the mobile phase from the first chromatographic column 110, the front impurity (a) is first discharged by the third multi-way valve 350; the front impurity and sample mixing section (b) is transferred to the second chromatographic column 120 through the third driving pump 430, and a small amount of qualified sample section (c) is transferred to the second chromatographic column 120 through the third driving pump 430; the rear impurity and sample mixing section (d ) is transferred to the second chromatographic column 120 through the third driving pump 430. At this time, the front impurity and sample mixing section (b) and a small amount of qualified sample section and the rear impurity and sample mixing section (d) together constitute the inner circulation sample (f). During the transfer of the inner circulation sample (f), the entire system is controlled to be in a closed-loop state, and no additional elution solvent is required. At this stage, according to the application scenario of the sample, the fourth driving pump 440 is turned on to pump in the diluent for online dilution and the fifth driving pump 450 is turned on under the same conditions to perform the transfer. After the transfer is completed, the first control valve 310 is switched to the regeneration agent, and the first driving pump 410 is controlled to discharge the rear impurities (e) of the first chromatographic column 110; finally, the first control valve 310 is switched to the elution solvent, and the first driving pump 410 is controlled to balance the first chromatographic column 110, and the first cycle is completed.

[0090] At the beginning of the second cycle, the inner circulation sample (f) of the second chromatographic column 120 is transferred to the first chromatographic column 110 through the inlet of the first chromatographic column 110 for receiving the inner circulation sample (f). During the transfer of the inner circulation sample (f), the entire system is controlled to be in a closed-loop state without adding additional elution solvent. At this time, according to the sample application scenario, the fourth drive pump 440 is turned on to pump in the diluent for online dilution and the fifth drive pump 450 with the same flow rate is turned on at the same time for transfer. After the transfer is completed, the first control valve 310 is switched to the regeneration agent, and the first drive pump 410 is turned on to discharge the impurities (e) after the second chromatographic column 120; finally, the first control valve 310 is switched to the elution solvent, and the first drive pump 410 is turned on to balance the second chromatographic column 120, and the second cycle ends.

[0091] And, repeating the first cycle and the second cycle until the inner cycle sample (f) meets the quality requirements.

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

[0093] The developing parameters include: the first chromatographic column 110 and the second chromatographic column 120 are in an independent state, the developing reagent is ethanol with a volume fraction of 96%, the solvent pump flow rate of the developing reagent is 5 mL / min, and the developing time is 5 min.

[0094] The parameters for transferring the front impurity and sample mixing section, the parameters for transferring a small amount of qualified samples, and the parameters for transferring the rear impurity and sample mixing section all include: the first chromatographic column 110 and the second chromatographic column 120 are connected in series in a closed loop, the eluent is the solvent in the second chromatographic column 120, and the circulation pump flow rate is 5 mL / min; the parameters for transferring the internal circulation sample in the first chromatographic column 110 to the fourth driving pump 440 and the fifth driving pump 450 in the second chromatographic column 120 include: the flow rates of the fourth driving pump 440 and the fifth driving pump 450 are 0.2 mL / min, and the diluent is purified water.

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

[0096] At the beginning of the second cycle, the inner circulation sample of the second chromatographic column 120 is transferred to the first chromatographic column 110 through the inlet of the first chromatographic column 110 for receiving the inner circulation sample. During the transfer of the inner circulation sample, the entire system is in a closed-loop state and there is no need to add additional elution solvent. At this time, according to the sample application scenario, the fourth drive pump 440 is turned on for online dilution and the fifth drive pump 450 with the same flow rate is turned on for transfer. After the transfer is completed, the solvent selection valve V01 is switched to the regeneration agent and the P01 pump is turned on to discharge the impurities after the second chromatographic column 120; finally, the solvent selection valve V01 is switched to the elution solvent and the P01 pump is turned on to balance the second chromatographic column 120, and the second cycle ends.

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

[0098] The parameters for the internal circulation sample transfer include: the first chromatographic column 110 and the second chromatographic column 120 are connected in series in a closed loop, the eluent is the solvent in the first chromatographic column 110, and the circulation pump flow rate is 5 mL / min; the parameters for the internal circulation sample transfer from the second chromatographic column 120 to the fourth drive pump 440 and the fifth drive pump 450 in the first chromatographic 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 chromatographic column 110 and the second chromatographic column 120 are independent, the equilibration liquid and the developing reagent are the same, the regeneration liquid is pure ethanol, the flow rate of the first driving pump 410 of the eluent is 5 mL / min, and the regeneration and equilibration time are 3 minutes each.

[0100] After the sample has gone through four large cycles, a qualified sample is collected from the second valve port of the second chromatographic column 120 .

[0101] The purity of the collected qualified samples was determined by gas chromatography, and the gas phase conditions included: instrument model: Agilent GC8860, chromatographic 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 2.5℃ / min to 270℃, holding time 5min, FID detector: 270℃.

[0102] The obtained gas chromatographic detection graph is shown in FIG3 , which is a detection spectrum of the single-column and double-column alternating circulation chromatography system 10 for separation and purification of oil after separation and purification according to an embodiment of the present invention. As shown in FIG3 , FIG3 is a detection spectrum of qualified samples collected after preparation, and the raw materials are transferred through multiple cycles, and finally the sample and impurities are well separated, and the purity of qualified products can be greater than 99%, and the calculated yield is greater than 90%.

[0103] In summary, the present invention reduces the number of chromatographic columns and utilizes the countercurrent operation of a simulated moving bed SMB to achieve continuous and automated cyclic operation of a double-column chromatography. Its main advantage is that two chromatographic columns are used to achieve the functions of countercurrent operation and continuous operation of a simulated moving bed, and a multi-component mixture can be truly and continuously separated and purified at a lower investment cost, achieving the advantages of 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 the ability to significantly reduce the amount of eluent used.

[0104] In the above embodiments, the description of each embodiment has its own emphasis. For parts that are not described in detail in a certain embodiment, reference can be made to the relevant descriptions of other embodiments.

[0105] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described 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 above-mentioned embodiments only express several implementation methods of the present invention, and the description thereof is relatively specific and detailed, but it cannot be understood as limiting the scope of the patent of the present invention. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A single-column and double-column alternating circulation chromatography system for separation and purification, characterized in that: It 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-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, wherein 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 also 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 simultaneously 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 simultaneously connected to the first multi-way valve, and the fourth multi-way valve is also simultaneously connected to the first multi-way valve, the fifth multi-way valve and the sixth multi-way valve.

2. The single-column and double-column alternating circulation 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 independently selected from four-way valves.

3. The single-column and double-column alternating circulation chromatography system for separation and purification according to claim 1, characterized in that: The single-column and double-column alternating circulation chromatography system for separation and purification also includes a first driving pump. One of the valve ports of the first multi-way valve is connected in parallel with one of the valve ports of the second multi-way valve and then connected to the first control valve through the first driving pump.

4. The single-column and double-column alternating circulation chromatography system for separation and purification according to claim 1, characterized in that: The single-column and double-column alternating circulation chromatography system for separation and purification also includes a second driving pump. One of the valve ports of the first multi-way valve is connected in parallel with one of the valve ports of the second multi-way valve and then connected to the second control valve through the second driving pump.

5. The single-column and double-column alternating circulation chromatography system for separation and purification according to claim 1, characterized in that: The single-column and double-column alternating circulation chromatography system for separation and purification also includes a third driving pump. One of the valve ports of the fourth multi-way valve, one of the valve ports of the fifth multi-way valve, and one of the valve ports of the sixth multi-way valve are connected in parallel and connected to the first multi-way valve through the third driving pump.

6. The single-column and double-column alternating circulation chromatography system for separation and purification according to claim 1, characterized in that: The single-column and double-column alternating circulation chromatography system for separation and purification further comprises a fourth driving pump, which is connected to one of the valve ports of the fifth multi-way valve for pumping in a diluent; And / or, the single-column and double-column alternating circulation chromatography system for separation and purification further comprises a fifth driving pump, and the fifth driving pump is connected to one of the valve ports of the sixth multi-way valve for pumping out overflow solvent.

7. The single-column and double-column alternating circulation 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 an ultraviolet detector, an evaporative light detector, and a differential refractive index detector; And / or, the second detector includes one or more of an ultraviolet detector, an evaporative light detector and a differential refractive index detector.

8. The single-column and double-column alternating circulation chromatography system for separation and purification according to claim 1, characterized in that: The first multi-way valve at least comprises 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 driving pump; The second multi-way valve at least comprises 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 connected to both the fifth multi-way valve and the sixth multi-way valve; The third multi-way valve at least includes a first valve port connected to the first detector and used to discharge the front impurities, a second valve port connected to the second multi-way valve and the fifth multi-way valve, a third valve port used to discharge the rear impurities, and a fourth valve port used to discharge the separated sample; The fourth multi-way valve at least includes a first valve port connected to the second detector and used to discharge the residual impurities, a second valve port used to discharge the residual impurities, a third valve port used to discharge the separated sample, and a second valve port connected to the third driving pump, the fifth multi-way valve and the sixth multi-way valve at the same time; The fifth multi-way valve at least comprises a first valve port, a second valve port connected to the second multi-way valve, a third valve port for pumping in a diluent, and a fourth valve port connected to both the first multi-way valve and the fourth multi-way valve; The sixth multi-way valve at least includes 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 multi-way valve and the fourth multi-way valve.

9. A single-column and double-column alternating cycle chromatography method for separation and purification, characterized in that: The single-column and double-column alternating circulation chromatography system for separation and purification according to any one of claims 1 to 8 comprises the following steps: In a first cycle, the multi-component mixture (G) is saturatedly loaded through the first chromatographic column, and the first chromatographic column is eluted. The multi-component mixture (G) is layered in the first chromatographic column and sequentially divided into a pre-impurity (a), a pre-impurity and sample mixed section (b), a small amount of qualified sample section (c), a post-impurity and sample mixed section (d), and a post-impurity section (e); under the elution of the first chromatographic column with the mobile phase, the pre-impurity (a) is first discharged by the third multi-way valve; the pre-impurity and sample mixed section (b) is transferred to the second chromatographic column, and the small amount of qualified sample section (c) is transferred to the The second chromatographic column; the post-impurity and sample mixing section (d) is transferred to the second chromatographic column, and the pre-impurity and sample mixing section (b), a small amount of qualified sample section and the post-impurity and sample mixing section (d) together constitute the inner circulation sample (f). During the transfer of the inner circulation sample (f), the entire system is controlled to be in a closed loop state, and no additional elution solvent is required. At this stage, a diluent is pumped in for online dilution according to the application scenario of the sample; the first control valve is switched to a regenerant to discharge the post-impurity (e) of the first chromatographic column; the first control valve is switched to an elution solvent to balance the first chromatographic column, and the first cycle ends; The second cycle begins, the inner circulation sample (f) of the second chromatographic column is transferred to the first chromatographic column through the inlet of the first chromatographic column for receiving the inner circulation sample (f), and the whole system is controlled to be in a closed loop state during the transfer of the inner circulation sample (f), and no additional elution solvent is needed. At this time, a diluent is pumped in for online dilution according to the sample application scenario; the first control valve is switched to a regeneration agent, and the first drive pump is turned on to discharge the impurities (e) after the second chromatographic column; the first control valve is switched to an elution solvent, and the first drive pump is turned on to balance the second chromatographic column, and the second cycle ends; And, repeating the first cycle and the second cycle until the inner cycle sample (f) meets the quality requirements.

10. The single-column and double-column alternating cycle chromatography method for separation and purification according to claim 9, characterized in that: The single-column and double-column alternating cycle chromatography method for separation and purification also satisfies at least one of the following conditions: (1) collecting and storing the front impurities (a) or the rear impurities (e) at the outer discharge of the first chromatographic column and the second chromatographic column respectively; (2) The mobile phase includes but is not limited to the eluent, regeneration solution or equilibrium solution; (3) After repeating the first cycle and the second cycle at least once, regenerating or balancing the first chromatographic column and / or the second chromatographic column by pumping in a regeneration liquid and a balancing liquid through a first driving pump.

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