Modulator

Abstract

The present invention relates to a modulator. The modulator comprises: a first two-way multi-port valve, a dilution pump, a collector, a second-dimensional chromatographic column, a second-dimensional gradient pump and a modulation controller. The first two-way multi-port valve has a first port, a second port and a third port, the first port is used to receive sample fractions from a conventional liquid chromatography system, and the second port is used to communicate with a detector of the conventional liquid chromatography system. The collector is used to collect sample fractions from the third port. The second-dimensional chromatographic column is used to separate the sample fractions from the collector. The second-dimensional gradient pump is used to inject a second-dimensional mobile phase into the second-dimensional chromatographic column. The modulation controller is used to control the first two-way multi-port valve and the second-dimensional gradient pump respectively. Since the above-mentioned modulator can be independently controlled by the modulation controller to perform sampling, injection and second-dimensional liquid chromatography separation, the above-mentioned modulator combined with various conventional liquid chromatography systems can constitute a two-dimensional liquid chromatography system.

Description

Technical Field

[0001] The invention relates to the technical field of liquid chromatography separation systems, in particular to a modulator. Background Art

[0002] Commercial two-dimensional liquid chromatography systems mostly use a solution of joint control of one-dimensional, two-dimensional, detectors and switching valves. The system has a high degree of integration, which can ensure the coordinated control of the modulation process and the integration of data acquisition. However, this integrated design also makes the two-dimensional liquid chromatography system expensive. In addition, it can only be used for a dedicated machine and cannot realize two-dimensional chromatography construction by combining existing equipment from different manufacturers. It is difficult for ordinary users to afford it. Summary of the invention

[0003] Based on this, it is necessary to provide a modulator that can utilize existing chromatography equipment, is easy to transplant and can reduce the cost of the two-dimensional chromatography system in order to address the high price of the traditional two-dimensional chromatography system.

[0004] The embodiment of the present application provides a modulator for connecting to a conventional liquid chromatography system, wherein the conventional liquid chromatography system comprises: a first-dimensional controller, a first-dimensional gradient pump, a first-dimensional injector, a first-dimensional chromatographic column, and a first-dimensional detector; the first-dimensional controller is used to control the first-dimensional gradient pump, the first-dimensional injector, and the first-dimensional detector respectively; the first-dimensional gradient pump is used to transport the first-dimensional mobile phase to the first-dimensional chromatographic column through the first-dimensional injector, so that the sample to be separated in the first-dimensional chromatographic column is separated under the action of the first-dimensional mobile phase, thereby obtaining the sample fraction separated by the first-dimensional chromatographic column;

[0005] The modulator comprises: a first two-position multi-way valve, a collector, a second-dimension chromatographic column, a second-dimension gradient pump and a modulation controller;

[0006] The first two-position multi-way valve has a first port, a second port, a third port and a fourth port, the first port is used to communicate with the outlet end of the first-dimension chromatographic column to receive the sample fraction from the first-dimension chromatographic column, and the second port is used to communicate with the first-dimension detector;

[0007] The first two-position multi-way valve has a first state and a second state, in which the first port is communicated with the second port; in the second state, the first port is communicated with the third port, so that the sample fraction flows from the first port to the third port, and the outlet end of the second-dimensional chromatographic column is connected to the fourth port, and the fourth port is communicated with the second port;

[0008] The collector is used to collect the sample fraction from the third port;

[0009] The second-dimensional chromatographic column is used to separate the sample fractions from the collector;

[0010] The second dimension gradient pump is used to inject the second dimension mobile phase into the second dimension chromatographic column;

[0011] The modulation controller is used to control the first two-dimensional multi-way valve and the second-dimensional gradient pump respectively.

[0012] The inlet and outlet ends of the modulator are connected to a conventional liquid chromatography system respectively, so as to form a two-dimensional liquid chromatography system. Since the modulator can be independently controlled by an independent modulation controller to perform sampling, injection and second-dimensional liquid chromatography separation, there is no technical barrier when the modulator is combined with conventional liquid chromatography systems of various manufacturers or brands. In this way, the detection, data acquisition and analysis functions of the conventional liquid chromatography system can be fully utilized without independent configuration. The two-dimensional liquid chromatography system can be combined with a large number of conventional liquid chromatography systems configured in conventional analytical laboratories to construct a large-scale conventional liquid chromatography system. Since a large number of existing conventional liquid chromatography systems and low-cost modulators are combined, the cost of the two-dimensional liquid chromatography can be greatly reduced and its flexibility of use can be improved, which is convenient for the promotion and application of the two-dimensional liquid chromatography system.

[0013] In one embodiment, two collectors are provided between the first two-position multi-way valve and the second-dimensional chromatographic column, wherein one of the collectors is connected to the third port for collecting the sample fractions from the third port, and the other collector is connected to the second-dimensional chromatographic column for transferring the sample fractions into the second-dimensional chromatographic column; wherein the two collectors alternately collect the sample fractions from the third port and alternately transfer the sample fractions into the second-dimensional chromatographic column.

[0014] In one embodiment, the modulator also includes a switching valve controlled by the modulation controller, and the collector is connected to the switching valve; the switching valve can switch the state so that the two collectors alternately collect the sample fractions from the third port and alternately transfer the sample fractions to the second-dimensional chromatographic column.

[0015] In one embodiment, a plurality of collectors are provided between the first two-position multi-way valve and the second-dimensional chromatographic column, and the plurality of collectors are alternately connected to the third port in sequence to alternately collect the plurality of sample fractions from the third port in sequence, wherein the collectors correspond one-to-one to the sample fractions; after the plurality of collectors respectively complete the collection of the corresponding sample fractions, the plurality of collectors are alternately connected to the second-dimensional chromatographic column in sequence to alternately transfer the plurality of sample fractions into the second-dimensional chromatographic column in sequence.

[0016] In one embodiment, the modulation also includes a switching valve controlled by the modulation controller, and the collector is connected to the switching valve; the switching valve can switch the state, so that multiple collectors collect the sample fractions from the third port in turn and alternately or transfer multiple sample fractions to the second-dimensional chromatographic column in turn and alternately. The switching valve can switch the state.

[0017] In one embodiment, the switching valve is a two-position eight-way valve or a two-position ten-way valve or two connected two-position six-way valves; or

[0018] The switching valve is composed of a two-position ten-way valve, two two-position six-way valves respectively connected to the two-position ten-way valve, and a two-position four-way valve connected to the two-position ten-way valve.

[0019] In one embodiment, the modulator further comprises a dilution pump controlled by the modulation controller, wherein the dilution pump is connected to one end of the collector connected to the third port, so that the dilution solvent pumped out by the dilution pump merges with the sample fraction from the third port and enters the collector.

[0020] In one embodiment, the modulator is a second-dimensional injector controlled by the modulation controller, and the second-dimensional injector is connected to one end of the collector to inject the driving mobile phase into the collector, and the other end of the collector is connected to the second-dimensional chromatographic column to transfer the driving mobile phase and the sample fraction into the second-dimensional chromatographic column; the second-dimensional gradient pump is connected to one end of the second-dimensional chromatographic column connected to the collector.

[0021] In one embodiment, the first two-position multi-way valve is a two-position three-way valve or a two-position six-way valve.

[0022] In one embodiment, the collector is a quantitative loop or a trapping column. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is a schematic diagram of the connection relationship between the modulator of the first embodiment and a conventional liquid chromatography system;

[0024] Figure 2 for Figure 1 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the first two-position six-way valve is in the second state and the switching valve is in the first state;

[0025] Figure 3 for Figure 1 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the first two-position six-way valve is in the second state and the switching valve is in the second state;

[0026] Figure 4 It is a schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve of the second embodiment is in the first state;

[0027] Figure 5 for Figure 4 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the second state;

[0028] Figure 6 It is a schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve of the third embodiment is in the first state;

[0029] Figure 7 for Figure 6 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the second state;

[0030] Figure 8 It is a schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve of the fourth embodiment is in the first state;

[0031] Fig. 9 for Figure 8 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the second state;

[0032] Fig.10 for Figure 8 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the third state;

[0033] Fig.11 for Figure 8 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the fourth state;

[0034] Fig.12 for Figure 8 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the fifth state;

[0035] Fig.13 for Figure 8 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the sixth state;

[0036] Fig.14 for Figure 8 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the seventh state;

[0037] Fig.15 for Figure 8 A schematic diagram of the connection relationship between the modulator and the conventional liquid chromatography system when the switching valve is in the eighth state;

[0038] Fig.16 for Figure 8 A schematic diagram of a state in which the modulator in the embodiment and the conventional liquid chromatography system constitute a comprehensive two-dimensional liquid chromatography separation system;

[0039] Fig.17 for Fig.16 Schematic diagram of another state when the middle modulator and the conventional liquid chromatography system constitute a comprehensive two-dimensional liquid chromatography separation system. DETAILED DESCRIPTION

[0040] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to understand the disclosure of the present invention more thoroughly and comprehensively.

[0041] It should be noted that when a part is referred to as being "fixed to" another part, it may be directly on the other part or there may be a central part. When a part is considered to be "connected" to another part, it may be directly connected to the other part or there may be a central part at the same time. The terms "vertical", "horizontal", "left", "right" and similar expressions used herein are for illustrative purposes only and do not represent the only implementation method.

[0042] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. 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 related listed items.

[0043] Please refer to Figure 1 The first embodiment of the present application provides a modulator 100. The modulator 100 is used to be connected with a conventional liquid chromatography system 10 for combined use. The modulator 100 includes a first two-position multi-way valve 110, a collector 120, a second-dimensional chromatographic column 130, a second-dimensional gradient pump 140, and a modulation controller 150.

[0044] The conventional liquid chromatography system 10 is used for first-dimensional liquid chromatography separation. The conventional liquid chromatography system 10 includes a first-dimensional controller 11, a first-dimensional gradient pump 12, a first-dimensional injector 13, a first-dimensional chromatographic column 14, and a first-dimensional detector 15. The first-dimensional controller 11 controls the first-dimensional gradient pump 12, the first-dimensional injector 13, and the first-dimensional detector 15 respectively. The first-dimensional gradient pump 12 transports the first-dimensional mobile phase to the first-dimensional chromatographic column 14 through the first-dimensional injector 13, so that the sample to be separated in the first-dimensional chromatographic column 14 is separated under the action of the first-dimensional mobile phase, and then the sample fraction separated by the first-dimensional chromatographic column 14 is obtained.

[0045] The first two-position multi-way valve 110 has a first port 111, a second port 112 and a third port 113. The first port 111 is used to receive sample fractions from the conventional liquid chromatography system 10. The second port 112 is used to communicate with the first dimension detector 15 of the conventional liquid chromatography system 10. The first two-position multi-way valve 110 has a first state and a second state. Figure 1 In the first state, the first port 111 is connected to the second port 112. Figure 2 and Figure 3 In the second state, the first port 111 is connected to the third port 113 .

[0046] Specifically, in the present embodiment, the first two-position multi-way valve 110 is a two-position six-way valve. It is understandable that the first two-position multi-way valve 110 can also be a two-position three-way valve, etc. When the conventional liquid chromatography system 10 is used in combination with the modulator 100, the inlet of the modulator 100 is connected to the outlet of the conventional liquid chromatography system 10, so that the outlet end of the first dimension chromatographic column 14 is connected to the first port 111, so that the sample fraction separated by the first dimension chromatographic column 14 can enter the first port 111.

[0047] Please refer to Figure 2 and Figure 3 When the first two-position multi-way valve 110 is in the second state, the conventional liquid chromatography system 10 is connected to the modulator 100 to form a two-dimensional liquid chromatography system, and the first-dimensional component can enter the modulator for second-dimensional collection and separation. The third port 113 is connected to the first port 111, so that the sample fraction separated by the first-dimensional chromatographic column 14 flows from the first port 111 to the third port 113. One end of the collector 120 is connected to the third port 113, and the other end is connected to the second-dimensional chromatographic column 130, so that the sample fraction separated by the first-dimensional chromatographic column 14 flows from the third port 113 to the collector 120, and then transferred from the collector 120 to the second-dimensional chromatographic column 130.

[0048] The second-dimension gradient pump 140 is connected to the second-dimension chromatographic column 130. The second-dimension gradient pump 140 is used to pump out a second-dimension mobile phase different from the first-dimension mobile phase, and inject the second-dimension mobile phase into the second-dimension chromatographic column 130 to separate the sample fractions separated by the first-dimension chromatographic column 14. The outlet end of the second-dimension chromatographic column 130 can be connected to the inlet end of the first-dimension detector 15, so that the sample fractions separated by the second-dimension chromatographic column 130 can enter the first-dimension detector 15 for detection.

[0049] In this embodiment, one end of the collector 120 is connected to the second-dimensional gradient pump 140, and the other end is connected to the second-dimensional chromatographic column 130, so that the second-dimensional mobile phase of the second-dimensional gradient pump 140 can enter the collector 120, and then the second-dimensional mobile phase is mixed with the sample fraction in the collector 120 and enters the second-dimensional chromatographic column 130.

[0050] It can be understood that the first-dimensional mobile phase in the first-dimensional gradient pump 12 and the second-dimensional mobile phase in the second-dimensional gradient pump 140 both have concentration gradients, and the corresponding elution procedure is gradient elution.

[0051] The modulation controller 150 may be an industrial computer (IPC), a PLC controller, a single chip microcomputer, etc. The modulation controller 150 is used to control the switching of the first two-position multi-way valve 110 between the first state and the second state and the working state of the second-dimensional gradient pump 140 .

[0052] Specifically, when the first dimension controller 11 in the conventional liquid phase system 10 is started, the start of the modulation controller 150 is triggered so that the modulation controller 150 controls the second dimension gradient pump 140. The modulation controller 150 can control the start and stop of the second dimension gradient pump 140. The modulation controller 150 can also control the flow rate of the second dimension mobile phase pumped by the second dimension gradient pump 140. The modulation controller 150 can control the first two-position multi-way valve 110 to switch between the first state and the second state.

[0053] It is understandable that in other embodiments, the modulation controller 150 can also be started individually by manual operation.

[0054] Please refer to Figure 1 When the first two-position multi-way valve 110 is switched to the first state, the first port 111 is connected to the second port 112, and the conventional liquid chromatography system 10 constitutes a one-dimensional liquid chromatography system. The sample fraction separated by the first-dimensional chromatographic column 14 directly enters the first-dimensional detector 15 from the first port 111 through the second port 112 for detection.

[0055] The above-mentioned modulator 100 can be used in combination with a conventional liquid chromatography system 10. When the first two-position multi-way valve 110 is in the second state, the sample fraction separated by the first dimension chromatographic column 14 can be collected by the collector 120, and then the sample fraction collected by the collector 120 is transferred to the second dimension chromatographic column 130. The second dimension mobile phase pumped out by the second dimension gradient pump 140 is injected into the second dimension chromatographic column 130 to separate the sample fraction. The sample fraction separated by the second dimension chromatographic column 130 can enter the first dimension detector 15 for detection. When the first two-position multi-way valve 110 is switched to the first state, the conventional liquid chromatography system 10 can also be used only as a one-dimensional liquid chromatography system for first dimension separation. By controlling the state switching of the first two-position multi-way valve 110, one-dimensional liquid chromatography separation or two-dimensional liquid chromatography separation can be selected.

[0056] The inlet and outlet ends of the modulator 100 are respectively connected to a conventional liquid chromatography system 10 to form a two-dimensional liquid chromatography system. Since the modulator 100 can be independently controlled by the modulation controller 150 to perform sampling, injection, and second-dimensional liquid chromatography separation, there is no technical barrier when the modulator 100 is combined with conventional liquid chromatography systems 10 of various manufacturers or brands, and thus the modulator 100 can be combined with conventional liquid chromatography systems 10 of various manufacturers or brands to form a two-dimensional liquid chromatography system, which is convenient for promoting the two-dimensional liquid chromatography system. By combining the low-cost modulator 100 with the conventional liquid chromatography system 10 that is widely configured in conventional laboratories, a two-dimensional liquid chromatography system can be obtained with a relatively low investment.

[0057] Please refer to Figure 2 and Figure 3 In one embodiment, two collectors 120 are provided between the first two-position multi-way valve 110 and the second dimension chromatographic column 130. One of the collectors 120 is connected to the third port 113 to collect the sample fraction from the third port 113, and the other collector 120 is connected to the second dimension chromatographic column 130 to transfer the sample fraction to the second dimension chromatographic column 130. The two collectors 120 alternately collect the sample fraction from the third port 113 and alternately transfer the sample fraction to the second dimension chromatographic column 130.

[0058] In this embodiment, the modulator 100 further includes a switching valve 160 controlled by the modulation controller 150. The collector 120 is in communication with the switching valve 160. The switching valve 160 can switch states so that the collector 120 alternately collects sample fractions from the third port 113 and transfers sample fractions to the second dimension chromatographic column 130.

[0059] Specifically, Figure 2 and Figure 3As shown, the switching valve 160 is a two-position ten-way valve. It is understood that the switching valve 160 can also be a two-position eight-way valve. The two collectors 120 provided between the first two-position multi-way valve 110 and the second dimension chromatographic column 130 are respectively the first collector and the second collector. The process of the collector 120 collecting the sample fraction is called sampling. The process of the collector 120 transferring the sample fraction to the second dimension chromatographic column 130 is called injection. The switching valve 160 has a first state and a second state.

[0060] When the switching valve 160 is in the first state, the first collector samples and the second collector 120 injects samples. Specifically, when the switching valve 160 is in the first state, the first collector is connected to the third port 113, so that the sample fraction separated by the first-dimensional chromatographic column 14 enters the first collector from the third port 113, so that it is collected by the first collector; one end of the second collector is connected to the second-dimensional gradient pump 140, and the other end is connected to the second-dimensional chromatographic column 130, so that the second-dimensional mobile phase pumped out by the second-dimensional gradient pump 140 enters the second collector, and enters the second-dimensional chromatographic column 130 together with the sample fraction in the second collector, so as to perform second-dimensional separation.

[0061] When the switching valve 160 is in the second state, the first collector injects samples and the second collector 120 samples. Specifically, when the switching valve 160 is in the second state, the second collector 120 is connected to the third port 113, so that the sample fraction separated by the first-dimensional chromatographic column 14 enters the second collector 120 from the third port 113, so that it is collected by the second collector 120; one end of the first collector is connected to the second-dimensional gradient pump 140, and the other end is connected to the second-dimensional chromatographic column 130, so that the second-dimensional mobile phase pumped out by the second-dimensional gradient pump 140 enters the first collector, and enters the second-dimensional chromatographic column 130 together with the sample fraction in the first collector, so as to perform second-dimensional separation.

[0062] When the modulator 100 is working, the sampling timing of the collector 120 is precisely matched with the injection timing, so that the two collectors 120 work simultaneously and perform sampling and injection alternately, and the injection and sampling before the second-dimensional separation can be performed simultaneously, thereby improving the efficiency of the second-dimensional separation.

[0063] By modulating the controller 150 to control the switching state of the switching valve 160, the timing of the switching state of the switching valve 160 is matched with the sampling timing and injection timing of the collector 120. The switching valve 160 can realize the simultaneous operation of the two collectors 120 and alternately perform sampling and injection through the switching state, and then the injection and sampling before the second-dimensional separation can be performed simultaneously, thereby improving the efficiency of the second-dimensional separation.

[0064] Since the modulation controller 150 can independently program the switching valve and the second-dimensional gradient pump 140, the sampling time, injection time and the second-dimensional gradient cycle separation time of the collector 120 are highly synchronized, so that the sampling time, injection time and the second-dimensional gradient cycle separation time in each modulation cycle can be highly synchronized, and thus there is no technical barrier when the above modulator 100 is combined with conventional liquid chromatography systems 10 of various manufacturers or brands. By combining the low-cost modulator 100 with the conventional liquid chromatography system 10 that is widely configured in conventional laboratories, a two-dimensional liquid chromatography system can be obtained with a relatively low investment, which facilitates the promotion of the two-dimensional liquid chromatography system.

[0065] In one embodiment, the collector 120 is a quantitative loop or a capture column. Specifically in this embodiment, the collector 120 is a reverse capture column.

[0066] Please refer to Figure 2 and Figure 3 In one embodiment, the collector 120 is a reverse phase trapping column. The modulator 100 further includes a dilution pump 170 controlled by the modulation controller 150. The dilution pump 170 is connected to one end of the collector 120 connected to the third port 113, so that the dilution solvent pumped out by the dilution pump 170 merges with the sample fraction from the third port 113 and enters the collector 120.

[0067] Specifically, the sample fraction separated by the first dimension chromatographic column 14 enters the collector 120 through the third port 113. The dilution solvent pumped out by the dilution pump 170 enters the collector 120. Before entering the collector 120, the sample fraction separated by the first dimension chromatographic column 14 and the diluent are first merged in the pipeline and then enter the collector 120 together.

[0068] When a two-dimensional system of reverse phase chromatography and hydrophilic chromatography is constructed, the first-dimensional mobile phase is a water-rich mobile phase, and the dilution solvent pumped out by the dilution pump 170 is a high-water phase. The sample fraction separated by the first-dimensional chromatographic column 14 is diluted by the dilution solvent, and the water content is increased. Since the collector 120 is a reverse phase capture column, the water content of the sample to be entered into the reverse phase capture column is increased, and the organic phase is reduced, so that the target analyte in the sample fraction can be more efficiently captured by the reverse phase capture column.

[0069] Please refer to Figure 4 and Figure 5 The second embodiment of the present application provides a modulator 200. The structure of the modulator 200 of the second embodiment is basically the same as the structure of the modulator 100 of the first embodiment, and will not be repeated here. The following focuses on the differences between the modulator 200 of the second embodiment and the modulator 200 of the first embodiment.

[0070] The modulator 200 further includes a second dimension injector 280 controlled by the modulation controller 250. The second dimension injector 280 is connected to one end of the collector 220 to inject the driving mobile phase into the collector 220. The other end of the collector 220 is connected to the second dimension chromatographic column 230. The second dimension gradient pump 240 is connected to one end of the second dimension chromatographic column 230 connected to the collector 220 to inject the second dimension mobile phase into the second dimension chromatographic column 230.

[0071] Specifically, one end of the collector 220 is connected to the second dimension injector 280, and the other end is connected to the second dimension chromatographic column 230. The driving mobile phase pumped out by the second dimension injector 280 drives the sample fraction in the collector 220 to flow toward the second dimension chromatographic column 230. The second dimension mobile phase pumped out by the second dimension gradient pump 240 flows toward the second dimension chromatographic column 230. Driven by the driving mobile phase, the sample fraction merges with the second dimension mobile phase and is separated in the second dimension chromatographic column 230.

[0072] Since the sample fraction in the collector 220 carries the first-dimensional mobile phase, when it is transferred to the second-dimensional chromatographic column 230 for separation, there will be a problem of incompatibility between the first-dimensional mobile phase and the second-dimensional mobile phase, resulting in chromatographic peak broadening. In this embodiment, the second-dimensional gradient pump 240 is arranged downstream of the collector 220, so that the elution flow path of the second-dimensional mobile phase no longer flows through the collector 220. The second-dimensional mobile phase and the sample fraction transferred in the collector 220 are first merged before entering the second-dimensional chromatographic column 230 to achieve online mixing and dilution, and then enter the second-dimensional chromatographic column 230 for separation. Since the second-dimensional gradient pump 240 can be controlled by the modulation controller 250, the flow rate of the second-dimensional mobile phase can be controlled by the second-dimensional gradient pump 240, so the sample fraction transferred by the collector 220 can be controllably diluted by the second-dimensional mobile phase before entering the second-dimensional chromatographic column 230, which is conducive to achieving column head focusing when entering the second-dimensional chromatographic column 230 for separation, thereby suppressing the peak broadening problem in the second-dimensional separation process.

[0073] Furthermore, by controlling the second-dimension injector 280 and the second-dimension gradient pump 240 through the modulation controller 250, the dilution multiple of the transferred fraction can be precisely adjusted over a wide range simply by changing the flow rate of the second-dimension injector 280 and the flow rate of the second-dimension gradient pump 240, without the need to replace the diversion component, which is more convenient for actual operation and further improves the separation effect.

[0074] In one embodiment, the modulator 200 further includes a mixer (not shown). The mixer is connected to the second dimension chromatographic column 230 and the collector 220 respectively. The mixer is connected to the second dimension gradient pump 240. The mixer is used to mix the second dimension mobile phase and the sample fraction from the collector 220.

[0075] Before the sample fraction enters the second-dimensional chromatographic column 230, it enters the mixer driven by the driving mobile phase, and the second-dimensional mobile phase pumped out by the second-dimensional gradient pump 240 enters the mixer, so that the sample fraction and the second-dimensional mobile phase are mixed in the mixer, which can make the second-dimensional mobile phase more controllably diluted, thereby facilitating the second-dimensional mobile phase to achieve column head focusing when entering the second-dimensional chromatographic column 230 for separation, thereby better suppressing the problem of peak broadening in the second-dimensional separation process.

[0076] It is understandable that in other embodiments, a mixer may not be provided, and the sample fraction and the second-dimensional mobile phase may be mixed directly in the pipeline before the second-dimensional chromatographic column 230 or mixed and diluted online in the second-dimensional chromatographic column 230 .

[0077] Please refer to Figure 6 and Figure 7 The third embodiment provides a modulator 300. The structure of the modulator 300 of the third embodiment is basically the same as that of the modulator 200 of the second embodiment, and will not be described in detail here. The following focuses on the differences between the modulator 300 of the third embodiment and the modulator 200 of the second embodiment.

[0078] The switching valve is two connected two-position six-way valves. The first collector 321 is connected to a two-position six-way valve 361. The second collector 322 is connected to another two-position six-way valve 362. Both two-position six-way valves can switch states. By simultaneously switching the states of the two two-position six-way valves, the switching valve can be switched between the first state and the second state, so that the two collectors can alternately collect the sample fractions from the third port 313 and alternately transfer the sample fractions to the second dimension chromatographic column 330.

[0079] Please refer to Figure 8 to Figure 15 The fourth embodiment provides a modulator 400. The structure of the modulator 400 of the fourth embodiment is basically the same as that of the first, second and third embodiments, and will not be described in detail here. The following focuses on the differences between the modulator 400 of the fourth embodiment and the modulators of the first, second and third embodiments.

[0080] A plurality of collectors are disposed between the first two-position multi-way valve 410 and the second-dimension chromatographic column 430. The plurality of collectors are sequentially and alternately connected to the third port 413 to sequentially and alternately collect a plurality of sample fractions from the third port 413, wherein the collectors correspond to the sample fractions one by one. After the plurality of collectors have completed collecting the corresponding sample fractions, the plurality of collectors are sequentially and alternately connected to the second-dimension chromatographic column 430 to sequentially and alternately transfer a plurality of sample fractions to the second-dimension chromatographic column 430.

[0081] The multiple collectors are connected to the switching valve, and the switching valve can switch the state so that the multiple collectors sequentially and alternately collect the sample fractions from the third port 413 or sequentially and alternately transfer multiple sample fractions to the second-dimension chromatography column 430 .

[0082] Specifically, the first dimension chromatographic column 44 separates the sample to be separated in sequence according to the change of polarity, so that the sample fraction separated by the first dimension chromatographic column 44 is separated into a plurality of fractions in sequence according to the change of polarity. When performing the center-cut two-dimensional separation, each collector collects a fraction. In this embodiment, four fractions are taken as an example, and the first fraction, the second fraction, the third fraction and the fourth fraction are separated in sequence by the first dimension chromatographic column 44. The number of collectors is four, namely the first collector 421, the second collector 422, the third collector 423 and the fourth collector 424.

[0083] In this embodiment, the switching valve is composed of a two-position ten-way valve 461, two two-position six-way valves respectively connected to the two-position ten-way valve 461, and a two-position four-way valve 462 connected to the two-position ten-way valve 461. The two two-position six-way valves are respectively a first two-position six-way valve 463 and a second two-position six-way valve 464.

[0084] The first collector 421 and the second collector 422 are in communication with the first two-position six-way valve 463. The third collector 423 and the fourth collector 424 are in communication with the second two-position six-way valve 464. The first two-position six-way valve 463 and the second two-position six-way valve 464 are in communication with the two-position ten-way valve 461, respectively. The first two-position six-way valve 463 can switch a state so that one of the first collector 421 or the second collector 422 is selectively in communication with the two-position ten-way valve 461. The second two-position six-way valve 464 can switch a state so that one of the third collector 423 or the fourth collector 424 is selectively in communication with the two-position ten-way valve 461.

[0085] The two-position ten-way valve 461 is connected to the third port 413 of the first two-position multi-way valve 410, so that the sample fractions separated by the first-dimension chromatographic column 44 enter the two-position ten-way valve 461 through the third port 413, so that the sample fractions can enter each collector alternately in sequence through the two-position ten-way valve 461 and the two two-position six-way valves for sampling.

[0086] The two-position four-way valve 462 is respectively connected to the two-position ten-way valve 461, the second dimension injector 480 and the second dimension chromatographic column 430. The two-position four-way valve 462 can switch the state so that the second dimension injector 480 is connected or disconnected with the two-position ten-way valve 461, and the second dimension chromatographic column 430 is connected or disconnected with the two-position ten-way valve 461. When the second dimension injector 480 and the second dimension chromatographic column 430 are respectively connected with the two-position ten-way valve 461, the second dimension injector 480 and the second dimension chromatographic column 430 can be connected with each collector alternately in sequence, so that each collector can be injected alternately in sequence, so as to perform second-dimensional separation on the sample fractions transferred from each collector to the second dimension chromatographic column 430.

[0087] The time when each fraction is separated from the first dimension chromatographic column 44 matches the sampling time of the corresponding collector. The time when the switching valve switches the state matches the sampling time of the collector. In this embodiment, the switching valve has at least eight states. Figures 8 to 11 When the switching valve is alternately switched between the first state, the second state, the third state, and the fourth state, the first fraction, the second fraction, the third fraction, and the fourth fraction are alternately separated from the first dimension chromatographic column 44, and the first collector 421, the second collector 422, the third collector 423, and the fourth collector 424 are sampled in sequence. Specifically, the second dimension injector 480 and the second dimension chromatographic column 430 are disconnected from the two-position ten-way valve 461, respectively, and each collector cannot perform injection at this time. The third port 413 is alternately connected to the first collector 421, the second collector 422, the third collector 423, and the fourth collector 424 through the ten-way valve and the two six-way valves, and the first fraction, the second fraction, the third fraction, and the fourth fraction are alternately separated from the first dimension chromatographic column 44, so that the first fraction is completely collected in the first collector 421, the second fraction is completely collected in the second collector 422, the third fraction is completely collected in the third collector 423, and the fourth fraction is completely collected in the fourth collector 424.

[0088] After multiple collectors have completed sampling, injection is performed. The timing of the switching valve switching state matches the injection time of the collector. Figures 12 to 15In this embodiment, when the switching valve switches between the fifth state, the sixth state, the seventh state, and the eighth state in sequence, the first collector 421, the second collector 422, the third collector 423, and the fourth collector 424 are alternately sampled in sequence, and the first fraction, the second fraction, the third fraction, and the fourth fraction are sequentially injected into the second-dimension chromatographic column 430 for separation. Specifically, the first-dimension chromatographic column 44 stops separating samples, and each collector cannot sample. The second-dimension injector 480 and the second-dimension chromatographic column 430 are respectively connected to the two-position ten-way valve 461 through the two-position four-way valve 462. By switching the states of the two-position ten-way valve 461 and the two two-position six-way valves, the second-dimension injector 480 and the second-dimension chromatographic column 430 are connected to the first collector 421, the second collector 422, the third collector 423 and the fourth collector 424 alternately in turn, so that the first collector 421, the second collector 422, the third collector 423 and the fourth collector 424 are injected in turn, and the first fraction, the second fraction, the third fraction and the fourth fraction flow out from the corresponding collectors in turn and are injected into the second-dimension chromatographic column 430 in turn for separation, thereby realizing the center-cutting two-dimensional chromatographic separation.

[0089] It can be understood that the number of collectors can also be other numbers, and the heart-cutting two-dimensional chromatographic separation of the other number of fractions can be performed.

[0090] In one embodiment, when the modulator 400 is combined with a conventional liquid chromatography system 40, comprehensive two-dimensional liquid chromatography separation can also be performed. Fig.16 and Fig.17 , the state of the switching valve can be switched so that one of the second collector 422 and the fourth collector 424 is connected to the third port 413, and the other is connected to the second dimension injection pump. The second collector 422 and the fourth collector 424 work simultaneously, and they are alternately connected to the third port 413 and alternately connected to the second dimension injection pump, so that comprehensive two-dimensional liquid chromatography separation can be performed.

[0091] The technical features of the above embodiments may be combined arbitrarily. To make the description concise, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0092] 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 modulator for connecting to a conventional liquid chromatography system, the conventional liquid chromatography system comprising: a first dimension controller, a first dimension gradient pump, a first dimension injector, a first dimension chromatographic column, and a first dimension detector; The first-dimension controller is used to control the first-dimension gradient pump, the first-dimension injector and the first-dimension detector respectively; the first-dimension gradient pump is used to transport the first-dimension mobile phase to the first-dimension chromatographic column through the first-dimension injector, so that the sample to be separated in the first-dimension chromatographic column is separated under the action of the first-dimension mobile phase, thereby obtaining the sample fraction separated by the first-dimension chromatographic column; Characterized in that the modulator comprises: a first two-position multi-way valve, a collector, a second-dimensional chromatographic column, a second-dimensional gradient pump and a modulation controller; The first two-position multi-way valve has a first port, a second port, a third port and a fourth port, the first port is used to communicate with the outlet end of the first-dimension chromatographic column to receive the sample fraction from the first-dimension chromatographic column, and the second port is used to communicate with the first-dimension detector; The first two-position multi-way valve has a first state and a second state, in which the first port is communicated with the second port; in the second state, the first port is communicated with the third port, so that the sample fraction flows from the first port to the third port, and the outlet end of the second-dimensional chromatographic column is connected to the fourth port, and the fourth port is communicated with the second port; The collector is used to collect the sample fraction from the third port; The second-dimensional chromatographic column is used to separate the sample fractions from the collector; The second dimension gradient pump is used to inject the second dimension mobile phase into the second dimension chromatographic column; The modulation controller is used to control the first two-dimensional multi-way valve and the second-dimensional gradient pump respectively.

2. The modulator according to claim 1, characterized in that Two collectors are provided between the first two-position multi-way valve and the second-dimensional chromatographic column, wherein one of the collectors is connected to the third port for collecting the sample fractions from the third port, and the other collector is connected to the second-dimensional chromatographic column for transferring the sample fractions into the second-dimensional chromatographic column; wherein the two collectors alternately collect the sample fractions from the third port and alternately transfer the sample fractions into the second-dimensional chromatographic column.

3. The modulator according to claim 2, characterized in that It also includes a switching valve controlled by the modulation controller, and the collector is connected to the switching valve; the switching valve can switch the state so that the two collectors alternately collect the sample fractions from the third port and alternately transfer the sample fractions to the second-dimensional chromatographic column.

4. The modulator according to claim 1, characterized in that A plurality of collectors are provided between the first two-position multi-way valve and the second-dimensional chromatographic column, and the plurality of collectors are alternately connected to the third port in sequence, so as to alternately collect the plurality of sample fractions from the third port in sequence, wherein the collectors correspond to the sample fractions one by one; after the plurality of collectors respectively complete the collection of the corresponding sample fractions, the plurality of collectors are alternately connected to the second-dimensional chromatographic column in sequence, so as to alternately transfer the plurality of sample fractions to the second-dimensional chromatographic column in sequence.

5. The modulator according to claim 4, characterized in that It also includes a switching valve controlled by the modulation controller, and the collector is connected to the switching valve; the switching valve can switch the state, so that multiple collectors collect the sample fractions from the third port in turn and alternately or transfer multiple sample fractions to the second-dimensional chromatographic column in turn and alternately. The switching valve can switch the state.

6. The modulator according to claim 3 or 5, characterized in that The switching valve is a two-position eight-way valve or a two-position ten-way valve or two connected two-position six-way valves; or The switching valve is composed of a two-position ten-way valve, two two-position six-way valves respectively connected to the two-position ten-way valve, and a two-position four-way valve connected to the two-position ten-way valve.

7. The modulator according to claim 1, characterized in that It also includes a dilution pump controlled by the modulation controller, and the dilution pump is connected to one end of the collector connected to the third port, so that the dilution solvent pumped out by the dilution pump merges with the sample fraction from the third port and then enters the collector.

8. The modulator according to claim 1, characterized in that It also includes a second-dimensional injector controlled by the modulation controller, the second-dimensional injector is connected to one end of the collector to inject a driving mobile phase into the collector, and the other end of the collector is connected to the second-dimensional chromatographic column to transfer the driving mobile phase and the sample fraction into the second-dimensional chromatographic column; The second-dimensional gradient pump is connected to one end of the second-dimensional chromatographic column connected to the collector.

9. The modulator according to claim 1, characterized in that The first two-position multi-way valve is a two-position three-way valve or a two-position six-way valve.

10. The modulator according to claim 1, characterized in that The collector is a quantitative loop or a trapping column.

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