Circulating device for high-speed counter-current chromatography

Abstract

The utility model discloses a circulating device for high-speed counter-current chromatography. The circulating device comprises a plurality of control valves, a plurality of capture rings, a mobile phase storage tank, a pump, a fraction collector, a counter-current chromatography host, a detector and a chromatography workstation, the mobile phase storage tank is communicated with the pump pipeline; the pump is communicated with the counter-current chromatography host through a pipeline; the counter-current chromatography host is communicated with the detector through a pipeline; the detector is connected with the chromatographic workstation computer; circulating control valves are arranged on a pipeline for communicating the mobile phase storage tank with the pump and a pipeline for communicating the capturing ring with the fraction collector; and the capture rings are respectively communicated with the detector and the fraction collector through capture control valves through opening and closing pipelines. By arranging a plurality of capture rings connected in parallel and correspondingly arranged state switching control valves, various substances with similar solubility / partition coefficients and isomer substances contained in natural products can be circulated so as to achieve effective separation, and the purity of difficult-to-separate substances obtained after the substances are separated by the device can reach 98% or above.

Description

Technical Field

[0001] This application relates to the technical field of separation and purification of natural products, and in particular to a circulation device for high-speed counter-current chromatography. Background Art

[0002] High-speed counter-current chromatography (HSCCC) is a new type of liquid-liquid partition chromatography technology that has developed rapidly in the past two decades. For the specific device structure, refer to Figure 1 , and the existing high-speed counter-current chromatography includes: a mobile phase storage tank, a pump, a fraction collector, a counter-current chromatography main unit, a detector, and a chromatography workstation. A pump is provided on the pipeline connecting the mobile phase storage tank and the counter-current chromatography main unit; a detector is provided on the pipeline connecting the counter-current chromatography main unit and the fraction collector; the detector is electrically connected to the chromatography workstation. The solvent in the mobile phase storage tank sends the separated sample into the counter-current chromatography main unit through the pump. After separation, each separated substance in the sample enters the detector for detection and the results are output through the chromatography workstation. The separated sample is collected by the fraction collector.

[0003] This method relies on the directionality of the polytetrafluoroethylene (PTFE) serpentine tube set in the counter-current chromatography main unit and the centrifugal force field generated by a specific high-speed planetary rotation, so that the stationary phase without carrier support can be stably retained in the serpentine tube, and the mobile phase can pass through the stationary phase unidirectionally and at a low speed. In a short time, the sample can be rapidly partitioned in the immiscible two-phase solvent system, and then the purpose of continuous counter-current extraction and separation of substances can be achieved.

[0004] The solute to be separated is partitioned between two immiscible phases, and the partition coefficient K of each solute is the ratio of the concentration of the solute in the stationary phase and the mobile phase. When K = 1, it means that the solute has the same solubility in both phases. K = 10 means that the solute has a higher solubility in the stationary phase than in the mobile phase and has a strong retention. K = 0 means that the solute is not partitioned in the stationary phase and will be eluted within one mobile phase volume.

[0005] Therefore, if the partition coefficients of several components are different, good separation can be achieved in HSCCC. On the contrary, if the partition coefficients of each component are similar, it is very difficult to achieve one-time separation. When the substance to be separated contains a large number of substances with the same polarity, especially isomers with similar structures, it is very difficult to separate the components with similar polarities or isomers at one time using pure high-speed counter-current chromatography.

[0006] The solution to such problems in the prior art is to propose specific mobile phases, stationary phases, and operating methods for specific substances to be separated. For example, the method for separating cis-trans isomers of sertraline hydrochloride by high-speed counter-current chromatography disclosed in CN201910041453.1. However, there are many types of substances to be separated in the prior art, and such methods have strong specificity and cannot be applied to the separation of other substances.

[0007] The information disclosed in the background art section is only intended to enhance the overall understanding of the present utility model and should not be regarded as an admission or any form of implication that such information constitutes the prior art already known to those of ordinary skill in the art. Summary of the Utility Model

[0008] The present application provides a recycling device for high-speed counter-current chromatography. This device can perform multiple segmented captures, conduct multiple cycles according to the needs of the separated substances, and achieve efficient separation of multiple substances with similar solubilities and isomers. The purity of the substances obtained after separation by this device can reach over 98%.

[0009] The present application provides a recycling device for high-speed counter-current chromatography, including: multiple control valves, multiple capture loops, a mobile phase storage tank, a pump, a fraction collector, a counter-current chromatography mainframe, a detector, and a chromatography workstation;

[0010] The mobile phase storage tank is connected to the pump through a pipeline; the pump is connected to the counter-current chromatography mainframe through a pipeline; the counter-current chromatography mainframe is connected to the detector through a pipeline; the detector is connected to the chromatography workstation computer;

[0011] A circulation control valve is provided on the pipeline connecting the mobile phase storage tank and the pump, as well as on the pipeline connecting the capture loop and the fraction collector;

[0012] Each capture loop is respectively connected to the detector and the pipeline for opening and closing the fraction collector through a capture control valve.

[0013] Preferably, the capture control valve is a capture four-way valve, including: position 1, position 2, position 3, and position 4; position 1, position 2, position 3, and position 4 are opened and closed for connection;

[0014] The circulation control valve is a circulation four-way valve, including: position 1, position 2, position 3, and position 4; position 1, position 2, position 3, and position 4 are opened and closed for connection.

[0015] Preferably, when performing cyclic capture, position 2 and position 3 of the circulation four-way valve are connected; for at least one capture loop, position 1 and position 4 of the capture four-way valve are connected, and position 2 and position 3 are connected; the capture loop is connected to the detector and the pump through pipelines.

[0016] Preferably, during separation, position 1 and position 2 of the circulation four-way valve are connected and communicated, and position 3 and position 4 are connected and communicated; the capture four-way valve of at least one capture loop is connected and communicated with the fraction collector pipeline.

[0017] Preferably, position 1 of the circulation four-way valve is connected and communicated with the mobile phase storage tank pipeline.

[0018] Preferably, position 2 of the circulation four-way valve is connected and communicated with the pump pipeline.

[0019] Preferably, position 3 of the circulation four-way valve is connected and communicated with the capture four-way valve pipeline of at least one capture loop.

[0020] Preferably, position 4 of the circulation four-way valve is connected and communicated with the fraction collector pipeline.

[0021] Preferably, the capture loop includes: a first capture loop, a second capture loop, a third capture loop to an Nth capture loop; the first capture loop, the second capture loop, the third capture loop to the Nth capture loop are in parallel; and are connected to the detector, the fraction collector, and the pump.

[0022] Preferably, the capture control valve is a capture four-way valve; the capture four-way valve includes: a second four-way valve, a third four-way valve, a fourth four-way valve to an nth four-way valve;

[0023] The second four-way valve is connected and communicated with the first capture loop, and switches the capture cycle state and the separation state through the second four-way valve;

[0024] The third four-way valve is connected and communicated with the second capture loop, and switches the capture cycle state and the separation state through the third four-way valve;

[0025] The fourth four-way valve is connected and communicated with the third capture loop, and switches the capture cycle state and the separation state through the fourth four-way valve;

[0026] The nth four-way valve is connected and communicated with the Nth capture loop, and switches the capture cycle state and the separation state through the nth four-way valve.

[0027] The beneficial effects that this application can produce include:

[0028] 1) The circulation device for high-speed countercurrent chromatography provided by this application, by setting multiple parallel capture loops and corresponding four-way valves for state switching, can effectively separate various substances with similar partition coefficients / solubilities and isomeric substances contained in natural products. The purity of the difficult-to-separate substances obtained after separating such substances by this device can reach more than 98%.

[0029] 2) The circulation device for high-speed countercurrent chromatography provided by this application has a simple structure. It is improved on the basis of high-speed countercurrent chromatography equipment and does not affect the accuracy of the separation and detection results of this equipment. Description of the Drawings

[0030] Figure 1 Schematic diagram of an existing high-speed countercurrent chromatography detection device;

[0031] Figure 2 Schematic diagram of a circulation device for high-speed countercurrent chromatography in at least one embodiment provided by the present application;

[0032] Figure 3 Schematic diagram of the state where the sample processed by the circulation device for high-speed countercurrent chromatography in at least one embodiment provided by the present application is not collected through the loop;

[0033] Figure 4 Schematic diagram of the state where the sample processed by the circulation device for high-speed countercurrent chromatography in at least one embodiment provided by the present application is collected through the loop;

[0034] Figure 5 Schematic diagram of the circulation state of the sample processed by the circulation device for high-speed countercurrent chromatography in at least one embodiment provided by the present application;

[0035] Figure 6 Schematic diagram of the four-way valve site in at least one embodiment provided by the present application;

[0036] Figure 7 Circulation countercurrent chromatogram obtained by processing natural substances in at least one embodiment provided by the present application;

[0037] Figure 8 For Figure 7 Liquid chromatogram of peak 2-1 in

[0038] Figure 9 For Figure 7 Liquid chromatogram of peak 2-2 in

[0039] Legend:

[0040] The first four-way valve 1, the second four-way valve 2, the third four-way valve 3, the fourth four-way valve 4, the first capture loop 11, the second capture loop 12, the third capture loop 13, the mobile phase storage tank 21, the pump 22, the fraction collector 211, the countercurrent chromatography main unit 221, the detector 222, the chromatography workstation 223. Detailed implementation manners

[0041] To make the objectives, technical solutions, and advantages of the embodiments of the present utility model clearer, the technical solutions in the embodiments of the present utility model will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present utility model. Obviously, the described embodiments are some, but not all, of the embodiments of the present utility model. Usually, the components of the embodiments of the present utility model described and illustrated herein can be arranged and designed in various different configurations.

[0042] Accordingly, the following detailed description of the embodiments of the present utility model provided in the drawings is not intended to limit the scope of the claimed present utility model, but merely represents selected embodiments of the present utility model. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present utility model without creative efforts fall within the scope of protection of the present utility model.

[0043] Technical means not detailed in this application and not used to solve the technical problems of this application are set according to common general knowledge in the art, and various common general knowledge setting methods can be implemented.

[0044] See Figures 2 to 6 , the circulation device for high-speed countercurrent chromatography provided in this application includes: a plurality of control valves, a plurality of capture loops, a mobile phase storage tank 21, a pump 22, a fraction collector 211, a countercurrent chromatography main unit 221, a detector 222, and a chromatography workstation 223;

[0045] The mobile phase storage tank 21 and the pump 22 are connected by a pipeline; the pump 22 and the countercurrent chromatography main unit 221 are connected by a pipeline; the countercurrent chromatography main unit 221 and the detector 222 are connected by a pipeline; the detector 222 and the chromatography workstation computer 223 are connected;

[0046] Circulation control valves are provided on the pipeline connecting the mobile phase storage tank 21 and the pump 22, and on the pipeline connecting the capture loop and the fraction collector 211.

[0047] Each capture loop is respectively connected to the pipeline of the detector 222 and the fraction collector 211 through a capture control valve for opening and closing.

[0048] The components not detailed therein are the same as those of the existing high-speed countercurrent chromatograph. The control valve can be a four-way or six-way valve; the braking method can be manual, pneumatic, or solenoid valve.

[0049] The device includes: a first four-way valve 1, a second four-way valve 2, a third four-way valve 3, a fourth four-way valve 4 to an Nth four-way valve; a first capture loop 11, a second capture loop 12, a third capture loop 13 to an Nth capture loop.

[0050] When performing conventional countercurrent chromatography separation, the 1st position and the 2nd position of the first four-way valve 1 are connected, the mobile phase storage tank 21 is connected to the 4th position of the first four-way valve 1, and the pump 22 is connected to the 3rd position of the first four-way valve 1. The pump 22 is connected to the countercurrent chromatography main unit 221 by a pipeline; the countercurrent chromatography main unit 221 and the detector 222 are connected by a pipeline; the detector 222 and the fraction collector 211 are connected by a pipeline. The detector 222 is electrically connected to the chromatography workstation 223. After the mobile phase separates the sample through the countercurrent chromatography main unit 221 and is detected by the detector 222, the obtained results are output through the chromatography workstation 223. The substances in the mobile phase enter the fraction collector 211 for collection after separation to obtain the substances.

[0051] When the obtained spectrogram shows that the substances to be separated are not effectively separated, such as Figure 7 the substance corresponding to peak 2 shown, the uneffectively separated components are first captured into the loop by opening the capture loop, and wait for cyclic separation. Figure 7 For substances with well-separated peaks shown in

[0052] When the first capture loop 11 captures the sample, the 1st position and the 2nd position of the first four-way valve 1 are connected, the mobile phase storage tank 21 communicates with the 4th position of the first four-way valve 1, and the pump 22 communicates with the 3rd position of the first four-way valve 1. After the 2nd position and the 3rd position of the second four-way valve 2 are connected, the detector 222 communicates with the 1st position of the second four-way valve 2; the first capture loop 11 communicates with the 4th position of the second four-way valve 2, so that the components to be separated enter the first capture loop 11. For the four-way valves arranged on the side of the capture loop that does not perform capture, the 1st position and the 2nd position are connected, the nth four-way valve is connected to the 3rd position pipeline of the first four-way valve 1, and the fraction collector is connected to the 4th position pipeline of the first four-way valve 1. When cycling is required, the 2nd position and the 3rd position of the first four-way valve 1 are connected. Under the push of the pump 22, substances that need to be separated multiple times can pass through the countercurrent chromatography main unit 221 multiple times to effectively separate substances with similar partition coefficients and isomers.

[0053] After the substances that have completed cycling are effectively separated, after the 1st position and the 2nd position of the first four-way valve 1 are connected, and after the 3rd position and the 4th position of the first four-way valve 1 are connected, the effectively separated substances flow out to the fraction collector 211 for collection.

[0054] Similarly, when the second capture loop 12 captures the sample, the second four-way valve 2 is closed to prevent the sample from entering the first capture loop 11. The 1st position and the 2nd position of the first four-way valve 1 are connected, the mobile phase storage tank 21 communicates with the 4th position of the first four-way valve 1, and the pump 22 communicates with the 3rd position of the first four-way valve 1; after the 2nd position and the 3rd position of the third four-way valve 2 are connected, the detector 222 communicates with the 1st position of the third four-way valve 2; the second capture loop 12 communicates with the 4th position of the third four-way valve 2, so that the components to be separated enter the second capture loop 12. For the four-way valves arranged on the side of the capture loop that does not perform capture, the 1st position and the 2nd position are connected, the nth four-way valve is connected to the 3rd position pipeline of the first four-way valve 1, and the fraction collector is connected to the 4th position pipeline of the first four-way valve 1; when cycling is required, the 2nd position and the 3rd position of the first four-way valve 1 are connected. Under the push of the pump 22, substances that need to be separated multiple times can pass through the countercurrent chromatography main unit 221 multiple times to effectively separate substances with similar partition coefficients and isomers.

[0055] After the substances completing the cycle are effectively separated, after the 1st position and the 2nd position of the first four-way valve 1 are connected, and after the 3rd position and the 4th position of the first four-way valve 1 are connected, the effectively separated substances flow out to the fraction collector 211 for collection.

[0056] When the third capture loop 13 captures the sample, the second four-way valve 2 and the third four-way valve 3 are closed to prevent the sample from entering the first capture loop 11 and the second capture loop 12. The 1st position and the 2nd position of the first four-way valve 1 are connected. The mobile phase storage tank 21 communicates with the 4th position of the first four-way valve 1, and the pump 22 communicates with the 3rd position of the first four-way valve 1. After the 2nd position and the 3rd position of the fourth four-way valve 2 are connected, the detector 222 communicates with the 1st position of the fourth four-way valve 2. The first capture loop 13 communicates with the 4th position of the third four-way valve 2, enabling the components to be separated to enter the first capture loop 13. The four-way valves provided on the side of the capture loop that is not performing capture all have the 1st position and the 2nd position connected. The nth four-way valve is connected to the 3rd position pipeline of the first four-way valve 1, and the fraction collector is connected to the 4th position pipeline of the first four-way valve 1. When circulation is required, the 2nd position and the 3rd position of the first four-way valve 1 are connected. Under the push of the pump 22, the substances that need to be separated multiple times can pass through the countercurrent chromatography main unit 221 multiple times to achieve effective separation of substances with similar distribution coefficients and isomers.

[0057] After the substances completing the cycle are effectively separated, after the 1st position and the 2nd position of the first four-way valve 1 are connected, and after the 3rd position and the 4th position of the first four-way valve 1 are connected, the effectively separated substances flow out to the fraction collector 211 for collection.

[0058] When the Nth capture loop captures the sample, the second four-way valve 2 to the (n - 1)th four-way valve are closed to prevent the sample from entering the first capture loop 11 to the (N - 1)th capture loop. The 1st position and the 2nd position of the first four-way valve 1 are connected. The mobile phase storage tank 21 communicates with the 4th position of the first four-way valve 1, and the pump 22 communicates with the 3rd position of the first four-way valve 1. After the 2nd position and the 3rd position of the nth four-way valve n are connected, the detector 222 communicates with the 1st position of the nth four-way valve 2. The (N - 1)th capture loop n communicates with the 4th position of the nth four-way valve 2, enabling the components to be separated to enter the (N - 1)th capture loop n. The four-way valves provided on the side of the capture loop that is not performing capture all have the 1st position and the 2nd position connected. The nth four-way valve is connected to the 3rd position pipeline of the first four-way valve 1, and the fraction collector is connected to the 4th position pipeline of the first four-way valve 1. When circulation is required, the 2nd position and the 3rd position of the first four-way valve 1 are connected. Under the push of the pump 22, the substances that need to be separated multiple times can pass through the countercurrent chromatography main unit 221 multiple times to achieve effective separation of substances with similar distribution coefficients and isomers.

[0059] After the substances completing the cycle are effectively separated, after the 1st position and the 2nd position of the first four-way valve 1 are connected, and after the 3rd position and the 4th position of the first four-way valve 1 are connected, the effectively separated substances flow out to the fraction collector 211 for collection.

[0060] The above device is applied to the actual separation of natural products, and the separation purity can reach more than 98%. For the specific treatment results, please refer to Figures 7 to 9 as shown. Figure 7 It is a cyclic countercurrent chromatogram. For the substances corresponding to peak 1, peak 3, peak 5, and peak 6 therein, high-purity compounds are obtained through high-speed countercurrent at one time. The compound corresponding to peak 2 cannot be obtained as a pure compound through high-speed countercurrent at one time. The above device is used to capture this component into the capture loop 11 first, and it passes through 5 cycles in the cyclic state, and finally the pure compounds peak 2-1 and peak 2-2 are separated. Figure 8 It is Figure 7 the liquid chromatogram of peak 2-1 in Figure 9 It is Figure 7 the liquid chromatogram of peak 2-2 in

[0061] Although the present invention has been described in detail with reference to the foregoing embodiments, for those skilled in the art, they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A circulation device for high speed countercurrent chromatography, characterized in that: include: A plurality of control valves, a plurality of capture loops, a mobile phase storage tank (21), a pump (22), a fraction collector (211), a countercurrent chromatography host (221), a detector (222), and a chromatography workstation (223); The mobile phase storage tank (21) and the pump (22) are connected by pipeline; the pump (22) and the countercurrent chromatography main unit (221) are connected by pipeline; the countercurrent chromatography main unit (221) and the detector (222) are connected by pipeline; the detector (222) and the chromatography workstation (223) are connected; A circulation control valve is provided on the pipeline connecting the mobile phase storage tank (21) and the pump (22), and on the pipeline connecting the capture loop and the fraction collector (211); Each capture ring is connected to the detector (222) and the open and closed pipeline of the fraction collector (211) through a capture control valve.

2. The circulation device for high speed countercurrent chromatography according to claim 1, characterized in that: The capture control valve is a capture four-way valve, including: position 1, position 2, position 3, and position 4; position 1, position 2, position 3, and position 4 are open and closed; The circulation control valve is a circulation four-way valve including: position 1, position 2, position 3, and position 4; position 1, position 2, position 3, and position 4 are open and closed.

3. The circulation device for high speed countercurrent chromatography according to claim 2, characterized in that: During cyclic capture, positions 2 and 3 of the circulating four-way valve are connected; positions 1 and 4, and positions 2 and 3 of the capture four-way valve of at least one capture ring are connected; and the capture ring is connected to the detector (222) and the pump (22) pipeline.

4. The circulation device for high speed countercurrent chromatography according to claim 3, characterized in that: During separation, positions 1 and 2 of the circulation four-way valve are connected, and positions 3 and 4 are connected; and the capture four-way valve of at least one capture ring is connected to the pipeline of the fraction collector (211).

5. The circulation device for high speed countercurrent chromatography according to claim 2, characterized in that: Position 1 of the circulation four-way valve is connected to a pipeline of the mobile phase storage tank (21).

6. The circulation device for high speed countercurrent chromatography according to claim 2, characterized in that: Position 2 of the circulation four-way valve is connected to the pump (22) pipeline.

7. The circulation device for high speed countercurrent chromatography according to claim 2, characterized in that: Position 3 of the circulation four-way valve is connected to a capture four-way valve pipeline of at least one capture ring.

8. The circulation device for high speed countercurrent chromatography according to claim 2, characterized in that: The circulating four-way valve is connected to the No. 4 fraction collector (211) pipeline.

9. The circulation device for high speed countercurrent chromatography according to claim 1, characterized in that: The capture rings include: a first capture ring (11), a second capture ring (12), a third capture ring (13) to an Nth capture ring; the first capture ring (11), the second capture ring (12), the third capture ring (13) to the Nth capture ring are connected in parallel; and are connected to a detector (222), a fraction collector (211), and a pump (22).

10. The circulation device for high speed countercurrent chromatography according to claim 9, characterized in that: The capture control valve is a capture four-way valve; the capture four-way valve comprises: a second four-way valve (2), a third four-way valve (3), a fourth four-way valve (4) to an nth four-way valve; The second four-way valve (2) is connected to the pipeline of the first capture ring (11), and the capture circulation state and the separation state are switched through the second four-way valve (2); The third four-way valve (3) is connected to the pipeline of the second capture ring (12), and the capture circulation state and the separation state are switched through the third four-way valve (3); The fourth four-way valve (4) is connected to the pipeline of the third capture ring (13), and the capture circulation state and the separation state are switched through the fourth four-way valve (4); The nth four-way valve is connected to the Nth capture loop pipeline, and the capture circulation state and the separation state are switched through the nth four-way valve.

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