A two-dimensional liquid chromatography separation system

Abstract

The present invention relates to the technical field of chromatographic preparation and separation. The purpose of the present invention is to provide a two-dimensional liquid chromatography separation system to solve the technical problem that the two-dimensional liquid chromatograph in the prior art has low efficiency and poor separation effect in separating different types of samples. The two-dimensional liquid chromatography separation system includes a one-dimensional chromatography system and a two-dimensional separation channel. The tail end of the one-dimensional chromatography system is provided with a distribution main valve, including an inlet and multiple outlets. There are at least two two-dimensional separation channels. Each two-dimensional separation channel includes a two-dimensional multi-position multi-way valve, a capture column, a two-dimensional pumping system, and a two-dimensional chromatography column. The two-dimensional multi-position multi-way valve includes a first inlet, a second inlet and multiple ports. The first inlet is connected to the outlet of the distribution main valve, the two-dimensional pumping system is connected to the second inlet, and the capture column and the two-dimensional chromatography column are connected to the corresponding ports; the two-dimensional separation channel has two working states, state one: the capture column is connected to the one-dimensional separation system separately; state two: the two-dimensional separation channel is self-operating.

Description

Technical Field

[0001] The invention relates to the technical field of chromatographic preparation and separation, and in particular to a two-dimensional liquid chromatography separation system. Background Art

[0002] Chromatography is a technical method based on the fact that different substances have different distribution coefficients in a system composed of a stationary phase and a mobile phase, and present different retention times during the elution process using the mobile phase, so that two mixed substances can be separated from each other. Chromatographic separation technology has developed into a variety of methods such as gas chromatography, liquid chromatography, supercritical fluid chromatography and electrochromatography. Liquid chromatography is a chromatographic technology that uses liquid as a mobile phase. Liquid chromatography has become one of the main means of separation analysis, purification and preparation at this stage due to its advantages such as high separation efficiency, high selectivity, high sensitivity, fast analysis speed, good repeatability and wide application range. It has been widely used in different fields such as chemistry, chemical engineering and medicine.

[0003] Liquid chromatography usually uses a complete set of liquid chromatography separation systems for separation operations. The most basic components of the liquid chromatography separation system include mobile phase containers, delivery pumps, injectors, chromatographic columns, collectors and other structures. The mobile phase and sample mixture are pumped into the chromatographic column by the delivery pump to achieve sample separation in the chromatographic column. In the prior art, the liquid chromatography separation system is also being gradually improved to meet the needs of multi-tube chromatographic columns and the separation of a large number of samples.

[0004] The Chinese invention patent with the authorization announcement number CN104330492B discloses a chromatographic separation device with intelligent automatic switching of multi-channel ion chromatographic columns, including multiple fluid channels. Different fluid channels are switched through a control panel so that samples in the fluid channels can flow through different ion chromatographic columns to separate the samples. Each ion chromatographic column is connected to the control panel in parallel through a chromatographic column connecting tube to form a fluid channel. This liquid chromatography separation device can separate a large number of samples through multiple chromatographic columns at the same time.

[0005] However, in current industrial needs and chemical processing, the composition of compound samples that need to be separated is becoming more and more complex, and the separation capacity of traditional liquid chromatography systems is close to the limit, and it is impossible to achieve a single complete separation. In order to automate, scale, and systematically separate and purify the sample components, it is necessary to perform two-dimensional separation on the fractions separated in the one-dimensional chromatographic column after the first separation. Therefore, the chromatographic separation system in the prior art is usually also connected to a two-dimensional liquid chromatography separation system at the rear end of the one-dimensional chromatographic system. The conventional two-dimensional liquid chromatography separation system has a simple structure, and a set of two-dimensional separation structures with the same or similar structure to the one-dimensional liquid chromatography system is directly connected to the rear end of the one-dimensional liquid chromatography separation system. The two-dimensional separation structure includes a two-dimensional chromatographic column, a detection and delivery pump, a collector and other structures. The two-dimensional separation structure is used to perform a secondary separation on the one-dimensional fraction in the one-dimensional liquid chromatography system, but the similarity with the one-dimensional separation is that the two-dimensional separation also has problems such as low efficiency and long separation time.

[0006] A Chinese invention patent with authorization announcement number CN106501429B discloses a multi-channel full two-dimensional liquid chromatograph with a multi-functional switching valve, including a one-dimensional chromatographic separation system and a multi-channel two-dimensional chromatographic separation system. The multi-channel two-dimensional separation system is adopted, and multiple chromatographic columns are used for two-dimensional separation. The one-dimensional chromatographic separation system and the multi-channel two-dimensional chromatographic separation system are connected through a multi-functional valve switching valve. The multi-functional switching valve includes an inlet connected to the rear end of the one-dimensional chromatographic separation system and an outlet connected to the inlet end of multiple two-dimensional chromatographic columns. The multi-functional switching valve is a one-inlet and multiple-outlet, multi-position and multi-channel selection valve. By switching the outlet of the multi-functional switching valve, different two-dimensional separation channels are switched to separate one-dimensional flow fractions.

[0007] However, the problem with the above two-dimensional liquid chromatograph when used is that, although each two-dimensional separation channel is connected in parallel to the outlet of the multifunctional switching valve so that samples can be separated in large quantities, this separation method is relatively simple and can only separate one type of sample. When changing samples, the one-dimensional chromatography system must be cleaned first and then the channel without injected flow fractions must be switched for sample separation. The whole process is cumbersome and cannot ensure that the two-dimensional separation channel with added samples and the cleaning fluid or another sample are mixed, and each two-dimensional separation channel cannot independently and efficiently separate different samples. There is no two-dimensional liquid chromatography device in the prior art that can be applied to the separation of large quantities of different types of samples. Summary of the invention

[0008] The object of the present invention is to provide a two-dimensional liquid chromatography separation system to solve the technical problems of low efficiency and poor separation effect of two-dimensional liquid chromatography in the prior art for separating different types of samples.

[0009] To achieve the above object, the technical solution of a two-dimensional liquid chromatography separation system provided by the present invention is:

[0010] A two-dimensional liquid chromatography separation system comprises a one-dimensional chromatography system and a two-dimensional separation channel. A distribution main valve is arranged at the tail end of the one-dimensional chromatography system. The distribution main valve is a multi-position multi-way valve. The distribution main valve comprises an inlet and a plurality of outlets. There are at least two two-dimensional separation channels. Each two-dimensional separation channel comprises a two-dimensional multi-position multi-way valve for communicating with each outlet of the distribution main valve. The system also comprises a capture column, a two-dimensional pumping system, and a two-dimensional chromatography column. The capture column is used to enrich the one-dimensional flow fraction. The two-dimensional chromatography column is used to separate the one-dimensional flow fraction enriched by the capture column. The two-dimensional multi-position multi-way valve comprises a first inlet, a second inlet, and a plurality of ports. The first inlet of the two-dimensional multi-position multi-way valve is connected to the outlet of the distribution main valve. The two-dimensional pumping system is connected to the second inlet. The trapping column and the two-dimensional chromatographic column are connected to the corresponding ports of the two-dimensional multi-position multi-way valve; the distribution main valve and the two-dimensional multi-position multi-way valve cooperate to realize two working states of the two-dimensional separation channel, working state one: the inlet of the distribution main valve is connected with the first inlet of the two-dimensional multi-position multi-way valve, and the one-dimensional chromatographic system is connected with the trapping column through the distribution main valve, the first inlet of the two-dimensional multi-position multi-way valve and the corresponding port, so as to realize the separate connection between the trapping column and the one-dimensional chromatographic system; working state two: the inlet of the distribution main valve is disconnected from the first inlet of the two-dimensional multi-position multi-way valve, the two-dimensional pumping system is connected with the trapping column through the second inlet of the two-dimensional multi-position multi-way valve and the corresponding port, and the trapping column is connected with the two-dimensional chromatographic column through the corresponding port, so as to realize the self-operation of the two-dimensional separation channel.

[0011] Beneficial effect: the one-dimensional fraction separated and eluted by the one-dimensional chromatography system can be introduced into each two-dimensional separation channel through the distribution main valve, and the two-dimensional multi-position multi-way valve at the front end of each two-dimensional separation channel can switch the connection state of the two-dimensional separation channel, so that the two-dimensional separation channel can receive the one-dimensional fraction in working state one and operate independently in working state two. When the two-dimensional separation channel is in working state two, the connection between the two-dimensional separation channel and the distribution main valve is disconnected, and the one-dimensional fraction enriched in the capture column is independently circulated to the two-dimensional chromatography column for separation through the two-dimensional pumping system, and will not interfere with or affect other two-dimensional separation systems, so it can be When replacing samples in the two-dimensional liquid chromatography system of the present invention, different samples can be separated simultaneously in the system; when replacing samples in the one-dimensional chromatography system, it is only necessary to switch the main distribution valve to disconnect the connection with each two-dimensional multi-position multi-way valve. Compared with the chromatography devices and systems in the prior art that are complicated and prone to cross-contamination, the two-dimensional liquid chromatography system in the present invention is independent, stable, and switches quickly, and has efficient reactions, thereby improving processing efficiency. It can be applied to the separation of large quantities and multiple types of samples, and effectively solves the technical problems of low efficiency and poor separation effect of two-dimensional liquid chromatographs in the prior art in separating different types of samples.

[0012] Preferably, the multiple ports of the two-dimensional multi-position multi-way valve include a capture column port and a chromatographic column port; the capture column ports are provided with two, including a first port and a second port, the first port is connected to one end of the capture column, the second port is connected to the other end of the capture column, and the chromatographic column port is connected to the two-dimensional chromatographic column; when the two-dimensional separation channel is in working state one, the first port is connected to the first inlet, and the capture column can enrich the one-dimensional flow fraction; when the two-dimensional separation channel is in working state two, the first port is connected to the chromatographic column port, and the second port is connected to the second inlet, so that the two-dimensional pumping system is connected to the capture column and the two-dimensional chromatographic column, so as to pump the one-dimensional flow fraction enriched by the capture column into the two-dimensional chromatographic column. The first port is connected to the first inlet as the liquid inlet of the capture column in working state one, and is connected to the two-dimensional chromatographic column as the liquid outlet of the capture column in working state two. The switching process is rapid, and no external pipeline or passage is required. The working state of the capture column can be directly changed. The structure is ingenious, so that the liquid phase can circulate quickly when the working state is switched.

[0013] Preferably, the multiple ports of the two-dimensional multi-position multi-way valve further include a waste discharge port, which is used to discharge waste liquid and impurities, and when the two-dimensional separation channel is in working state 1, the waste discharge port is connected to the second port. The waste discharge port can discharge impurities and waste liquid that are not captured by the capture column.

[0014] Preferably, the waste outlet is connected to a waste liquid pool, which collects impurities and waste liquid to prevent pollution.

[0015] Preferably, the two-dimensional separation channel also has a stop state. When the two-dimensional separation channel is in the stop state, the first inlet of the two-dimensional multi-position multi-way valve on the two-dimensional separation channel is disconnected from the outlet of the corresponding distribution main valve. At this time, the two-dimensional pumping system on the two-dimensional separation channel can be used to pump cleaning liquid to clean each channel component in the two-dimensional separation channel. When the two-dimensional separation channel is in the stop state, the one-dimensional chromatography system and the two-dimensional separation channel are disconnected. At this time, any operation can be performed on the two-dimensional separation channel, such as switching the connection state of the two-dimensional multi-position multi-way valve, replacing and repairing the channel component, or pumping cleaning liquid through the two-dimensional pumping system, so that the cleaning liquid flows through the connection route during self-operation to clean the two-dimensional separation channel.

[0016] Preferably, the two-dimensional separation channel further comprises a collection pipeline connected to the two-dimensional chromatographic column; the collection pipeline comprises a collector and a waste liquid pool connected in parallel to the rear end of the two-dimensional chromatographic column. The collection pipeline is set to collect the flow fractions in the two-dimensional chromatographic column to obtain the desired two-dimensional separation.

[0017] Preferably, the one-dimensional chromatography system comprises a one-dimensional multi-position multi-way valve and a one-dimensional chromatography column, wherein the one-dimensional multi-position multi-way valve comprises a plurality of inlets and an outlet connected to the front end of the one-dimensional chromatography column, wherein the inlet of the one-dimensional multi-position multi-way valve is respectively connected to an injector and a mobile phase reservoir, and the one-dimensional multi-position multi-way valve is used to introduce the initial sample and mobile phase in the injector into the one-dimensional chromatography column during transposition; the rear end of the one-dimensional chromatography column is connected to the inlet of the distribution main valve. The one-dimensional multi-position multi-way valve can also separate the injection process in the one-dimensional chromatography column, ensure the separation of the sample and the mobile phase before mixing, and prevent the sample or the mobile phase from being contaminated.

[0018] Preferably, a one-dimensional pumping system is connected between the front end of the one-dimensional chromatographic column and the outlet of the one-dimensional multi-position multi-way valve. The one-dimensional pumping system can ensure rapid one-dimensional separation operation.

[0019] Preferably, the one-dimensional chromatography system and each two-dimensional separation channel are provided with a detector, the one-dimensional detector of the one-dimensional chromatography system is connected between the one-dimensional chromatography column and the distribution main valve, and the two-dimensional detector of each two-dimensional chromatography column is connected to the rear end of the two-dimensional chromatography column. The detector monitors the capture or collection period of the flow fraction and detects the purity of the flow fraction, can determine whether the flow fraction is effectively separated and prepared, display the separation results in real time, provide judgment instructions and basis, and ensure accurate collection of the flow fraction.

[0020] Preferably, a mixer is provided between the one-dimensional chromatographic separation system and the inlet of the distribution main valve, and a regulating liquid pumping system is connected to the mixer. The regulating liquid pumping system can add regulating liquid to the one-dimensional fraction, and mix them through the mixer to adjust the polarity or pH value of the one-dimensional fraction, so that the capture column can effectively capture the one-dimensional fraction. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is a flow chart of the two-dimensional liquid chromatography separation system in Example 1 provided by the present invention;

[0022] Figure 2 for Figure 1 A schematic diagram of the structure of the left two-dimensional separation channel in the two-dimensional liquid chromatography separation system when it is in working state (the right two-dimensional separation channel is in a stopped state at this time);

[0023] Figure 3 for Figure 1 A structural schematic diagram of the left two-dimensional separation channel in the two-dimensional liquid chromatography separation system when it is in working state two (the right two-dimensional separation channel is in working state one at this time);

[0024] Figure 4 for Figure 1 Structural schematic diagram of the left two-dimensional separation channel and the right two-dimensional separation channel in the two-dimensional liquid chromatography separation system when both are in working state two.

[0025] Description of reference numerals:

[0026] 1. One-dimensional chromatography system; 2. Left two-dimensional separation channel; 3. Right two-dimensional separation channel; 4. Injector; 5. Mobile phase reservoir; 6. One-dimensional multi-position multi-way valve; 7. One-dimensional pumping system; 8. One-dimensional chromatographic column; 9. One-dimensional detector; 10. Mixer; 11. Regulating liquid pumping system; 12. Distribution main valve; 13. Two-dimensional multi-position multi-way valve; 14. Waste liquid tank; 15. Capture column; 16. Two-dimensional pumping system; 17. Two-dimensional chromatographic column; 18. Two-dimensional detector; 19. Collector; 20. First inlet; 21. Second inlet; 22. First port; 23. Second port; 24. Waste outlet; 25. Chromatographic column port. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical scheme and advantages of the present invention clearer, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention, that is, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. The components of the embodiments of the present invention described and shown in the drawings herein can be arranged and designed in various different configurations.

[0028] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without making creative work are within the scope of protection of the present invention.

[0029] It should be noted that relational terms such as "first" and "second" that may appear are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, terms such as "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device that includes a series of elements includes not only those elements, but also other elements that are not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, elements defined by the sentence "including a...", etc., do not exclude processes and methods that include the elements.

[0030] In the description of the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected" and "connected" that may appear 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 it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood by specific circumstances.

[0031] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the term "provided with" that may appear should be understood in a broad sense. For example, the object "provided with" may be a part of the body, or may be arranged separately from the body and connected to the body, and the connection may be a detachable connection or an inseparable connection. For those skilled in the art, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0032] The present invention is described in further detail below in conjunction with embodiments.

[0033] Specific embodiment 1 of the two-dimensional liquid chromatography separation system provided by the present invention:

[0034] like Figures 1 to 4 As shown, the two-dimensional liquid chromatography separation system includes a one-dimensional chromatography system 1 and two two-dimensional separation channels connected to the rear end of the one-dimensional chromatography system 1. Figures 1 to 4 The arrangement in the figure divides the two two-dimensional separation channels into a left two-dimensional separation channel 2 and a right two-dimensional separation channel 3, wherein the components of the two two-dimensional separation channels are the same, and both of the two two-dimensional separation channels have two working states, so that they can be connected or disconnected with the one-dimensional chromatography system 1 by switching the connection position, and can operate independently after switching the connection position.

[0035] like Figures 1 to 4As shown, the one-dimensional chromatography system 1 includes a one-dimensional multi-position multi-way valve 6 arranged at the front end, the one-dimensional multi-position multi-way valve 6 is a two-position three-way valve, including two inlets and one outlet, the one-dimensional multi-position multi-way valve 6 can switch two connecting positions, the two inlets are respectively connected with an injector 4 and a mobile phase reservoir 5, the injector 4 is used to store and pass the sample to be separated into the valve, the mobile phase reservoir 5 is used to store the mobile phase used by the one-dimensional chromatographic column 8, the mobile phase and the sample are passed into the one-dimensional chromatographic column 8 by switching the two connecting positions, and a one-dimensional pumping system 7 is also arranged between the front end of the one-dimensional chromatographic column 8 and the outlet of the one-dimensional multi-position multi-way valve 6, and the one-dimensional pumping system 7 pumps the mobile phase and the sample into the one-dimensional chromatographic column 8 respectively. A one-dimensional detector 9 is connected to the liquid outlet at the rear end of the one-dimensional chromatographic column 8, and the one-dimensional detector 9 is used to monitor the collection period of the one-dimensional flow fraction and detect the purity of the one-dimensional flow fraction. Among them, the rear end of the one-dimensional detector 9 is connected in series with a mixer 10 and a main distribution valve 12 in sequence. The mixer 10 is connected with a regulating liquid pumping system 11. The regulating liquid pumping system 11 is used to pump elution regulating liquid into the mixer 10 to mix with the one-dimensional flow fraction flowing out of the one-dimensional detector 9. The purpose of setting the mixer 10 is to make the one-dimensional flow fraction separated in the one-dimensional chromatographic column 8 and the elution regulating liquid pumped out of the regulating liquid pumping system 11 be fully mixed, so as to adjust the system polarity or pH value of the one-dimensional flow fraction obtained after the one-dimensional separation, so that the capture column can effectively capture the one-dimensional flow fraction and enrich the one-dimensional flow fraction.

[0036] like Figures 2 to 4 As shown, the distribution main valve 12 is a three-position four-way valve, including an inlet and three outlets. The inlet of the distribution main valve 12 is connected to the rear end of the mixer 10 to receive the one-dimensional fraction obtained in the one-dimensional chromatography system 1. The outlet of the distribution main valve 12 is used to connect the left and right two-dimensional separation channels, and there is another outlet used as a waste outlet 24, and the waste outlet 24 is connected to the waste liquid pool 14. The distribution main valve 12 can switch the communication lines therein so that its inlet is connected to different outlets, so that different two-dimensional separation channels or waste liquid pools 14 are connected to the one-dimensional chromatography system 1, so as to sequentially distribute the one-dimensional fraction of the one-dimensional chromatography system 1 to different two-dimensional separation channels or discharge impurities through the waste liquid pool 14 as needed.

[0037] like Figures 1 to 4As shown, the two two-dimensional separation channels each include a two-dimensional multi-position multi-way valve 13 connected to the outlet of the distribution main valve 12, and also include a capture column 15, a two-dimensional pumping system 16, a two-dimensional chromatographic column 17, a two-dimensional detector 18, and a two-dimensional collector 19 and a waste liquid pool connected in parallel (the two-dimensional collector 19 in the figure is replaced with a waste liquid pool when removing impurities), wherein the two-dimensional multi-position multi-way valve 13 includes a first inlet 20, a second inlet 21 and a plurality of ports, the ports include a capture column port, a chromatographic column port 25 and a waste outlet 24, the capture column 15 is connected to the respective channels. There are two column collection ports, including a first port 22 and a second port 23. The first port 22 is connected to one end of the capture column 15, and the second port 23 is connected to the other end of the capture column 15; the first inlet 20 of the two-dimensional multi-position multi-way valve 13 is connected to the outlet of the distribution main valve 12, the two-dimensional pumping system 16 is connected to the second inlet 21, the liquid inlet end of the two-dimensional chromatographic column 17 is connected to the chromatographic column port 25, the waste outlet 24 is connected to the waste liquid pool 14, and the liquid outlet end of the two-dimensional chromatographic column 17 is connected to the two-dimensional detector 18 and the two-dimensional collector 19 in sequence.

[0038] like Figures 2 to 4 As shown, the distribution main valve 12 and the two-dimensional multi-position multi-way valve 13 switch the connection passages therein correspondingly, so as to cooperate with each other to realize two working states of the two-dimensional separation channel. Working state one is that the distribution main valve 12 is connected to the first inlet 20 connected to the two-dimensional multi-position multi-way valve 13 through one of the outlets from the inlet, and the one-dimensional chromatography system 1 is connected to the capture column 15 through the distribution main valve 12, the first inlet 20 of the two-dimensional multi-position multi-way valve 13 and the first port 22, so that the capture column 15 in the two-dimensional separation channel is connected to the one-dimensional chromatography system 1 separately, and the capture column 15 can receive and enrich the one-dimensional flow fraction, and at this time, the liquid outlet end of the capture column 15 is connected to the second port 23, and the second port 23 is connected to the waste outlet 24 in the two-dimensional multi-position multi-way valve 13, so as to realize the discharge of impurities and waste liquid passing through the capture column 15. Working state one is Figure 2 The left two-dimensional separation channel 2 in the middle is in working state. At this time, the left two-dimensional separation channel 2 is connected and enriches the flow fraction in the one-dimensional chromatography system 1.

[0039] like Figure 3As shown, after one-dimensional fractions are enriched in the capture column 15 of the left two-dimensional separation channel 2, the connecting path in the distribution main valve 12 and the two-dimensional multi-position multi-way valve 13 can be switched to enter the second working state of the two-dimensional separation channel: the inlet of the distribution main valve 12 is disconnected from the first inlet 20 of the two-dimensional multi-position multi-way valve 13, the second inlet 21 of the two-dimensional multi-position multi-way valve 13 is connected to the second port 23, and the first port 22 is connected to the chromatographic column port. At this time, the two-dimensional pumping system 16 is connected to the capture column 15 and the two-dimensional chromatographic column 17. The two-dimensional pumping system 16 is started so that the one-dimensional fractions enriched in the capture column 15 can be pumped out. It is sent to the two-dimensional chromatographic column 17 to realize the self-operation of the two-dimensional separation channel. The two-dimensional pumping system 16 continuously pumps the two-dimensional mobile phase. The two-dimensional mobile phase drives the one-dimensional fraction to flow in the two-dimensional chromatographic column 17 and separate. During the separation process, the solution flowing out from the liquid outlet of the two-dimensional chromatographic column 17 enters the two-dimensional detector 18 and is detected by the two-dimensional detector 18 and its purity is displayed. According to the detection information of the two-dimensional detector 18, the two-dimensional collector 19 or the waste liquid pool 14 is selected to collect the two-dimensional fraction or discharge impurities and solvents. Finally, the target two-dimensional fraction can be collected according to the indication of the two-dimensional detector 18. It can be seen that when the left two-dimensional separation channel 2 enters the working state two, the left two-dimensional separation channel 2 self-operates, and the distribution main valve 12 is disconnected from the two-dimensional multi-position multi-way valve 13 of the left two-dimensional separation channel 2. At this time, the outlet of the distribution main valve 12 is connected to the first inlet 20 of the right two-dimensional separation channel 3, so that the right two-dimensional separation channel 3 enters the working state one. While the left two-dimensional separation channel 2 performs self-operating separation of one-dimensional flow fractions, the outlet of the distribution main valve 12 is switched to be connected to the right two-dimensional separation channel 3, so that the right two-dimensional separation channel 3 begins to enrich the one-dimensional flow fractions. The working operations of the left and right two-dimensional separation channels do not affect each other, and the working efficiency is greatly improved.

[0040] like Figure 4 As shown, after the trapping column 15 in the right two-dimensional separation channel 3 enriches the one-dimensional fraction, the connection between the distribution main valve 12 and the two-dimensional multi-position multi-way valve 13 in the right two-dimensional separation channel 3 is cut off, and the inlet and the port on the two-dimensional multi-position multi-way valve 13 are switched to make the two-dimensional pumping system 16, the trapping column 15 and the two-dimensional chromatographic column 17 in the two-dimensional separation channel interconnected. At this time, the left two-dimensional separation channel 2 and the right two-dimensional separation channel 3 are both in working state 2, and the two channels each separate the one-dimensional fraction enriched by the trapping column 15 through the two-dimensional chromatographic column 17 therein. Figure 4As shown, when both two-dimensional separation channels are in working state two, the inlet of the distribution main valve 12 is switched to be connected to the waste outlet 24, the one-dimensional chromatography system 1 is connected to the waste liquid pool 14, and the one-dimensional chromatography system 1 continues to run and enters the impurity removal state, and the impurities in the one-dimensional chromatography system 1 are further eluted and discharged, completing the regeneration of the one-dimensional chromatography column, and then the one-dimensional fraction preparation of the sample can be continued, or at this time, a strong eluent can be added to the one-dimensional chromatography system 1 to clean the components of the one-dimensional chromatography system 1.

[0041] Depend on Figures 2 to 3 The working state of the middle left two-dimensional separation channel 2 is switched or Figure 3 to Figure 4 It can be seen from the working state switching of the middle right two-dimensional separation channel 3 that when the two-dimensional separation channel is in working state one, the first port 22 connected to the capture column 15 is used as the liquid inlet of the capture column 15, and the second port 23 is used as the waste outlet of the capture column 15. At this time, the one-dimensional flow fraction enters the two-dimensional multi-position multi-way valve 13 from the first inlet 20 and enters the capture column 15 through the first port 22; when the two-dimensional separation channel is in working state two, the second port 23 is connected to the two-dimensional pumping system 16. At this time, the second port 23 is used as the liquid inlet of the capture column 15 to pump the mobile phase and pump pressure into the capture column 15, and the first port 22 is used as the liquid outlet of the capture column 15 to inject the one-dimensional flow fraction and the mobile phase enriched in the capture column 15 into the two-dimensional chromatographic column 17. In this embodiment, when the separation operation is completed, each two-dimensional separation channel is in a stopped state. At this time, the cleaning liquid can be pumped through the one-dimensional pumping system 7 or the regulating liquid pumping system 11 to pump the cleaning liquid to clean each component.

[0042] When the two-dimensional liquid chromatography separation system in this embodiment is used for separation operation, the one-dimensional chromatography system 1 can be first connected to the left two-dimensional separation channel 2, and the one-dimensional flow fraction can be enriched through the capture column 15 in the left two-dimensional separation channel. When the capture column 15 reaches the enrichment limit, the distribution main valve 12 can be directly switched to connect the one-dimensional chromatography system 1 to the right two-dimensional separation channel 3, and the capture column 15 in the right two-dimensional separation channel 3 continues to enrich the one-dimensional flow fraction; or the left two-dimensional separation channel 2 can be made to self-operate after the left two-dimensional separation channel 2 completes the enrichment, and then the distribution main valve 12 is connected to the waste liquid pool to regenerate the one-dimensional chromatography column and prepare new one-dimensional flow fractions, and then the distribution main valve 12 is switched to connect the one-dimensional chromatography system 1 to the right two-dimensional separation channel 3. Therefore, the two-dimensional liquid chromatography separation system in this embodiment can not only separate a single sample in large quantities, but also can replace samples or mobile phases when the left two-dimensional separation channel 2 switches to the working state 2 and self-operates to separate different samples.

[0043] In this embodiment, the one-dimensional pumping system 7 and the regulating liquid pumping system 11 are both medium-pressure isocratic elution pump systems composed of a single pump, and the internal flow rate is 1-100mL / min; each two-dimensional pumping system 16 is a high-pressure gradient elution pump system composed of two pumps, and the flow rate is 1-100mL / min; the one-dimensional chromatographic column 8 is a gel permeation chromatographic column in the prior art, with a specification of 50×410mm, and the two-dimensional chromatographic columns 17 are all C18 chromatographic columns with a specification of 30×250mm. The internal filler of the capture column 15 is C18 silica gel with a specification of 30×20mm.

[0044] The use process of the two-dimensional liquid chromatography separation system in this embodiment is as follows: taking tobacco crude extract as an example, nicotine extracted from tobacco is cyclically prepared, and the one-dimensional mobile phase used is an aqueous solution; the elution regulating liquid is an acetate buffer (pH=7.2), the two-dimensional mobile phase A is 15%-60% methanol, B is an acetate buffer (pH=7.2), and the separation elution gradient is: 0-60min, phase A 15%-60%. The process and method steps for extracting and separating nicotine from tobacco crude extract are as follows:

[0045] (1) preparing a sample and a mobile phase, crushing 500 g of tobacco raw material, sieving the powder through a sieve, adding 5000 mL of a 20% ethanol-water solution, heating at 60° C., reflux 3 times, each time for 2 hours, filtering to remove residues, and then filtering with a 50 μm filter membrane. After the filtrate is decompressed and recovered to remove the ethanol solvent, a tobacco crude extract is obtained;

[0046] (2) Add tobacco crude extract into the sample injector 4, add aqueous solution into the mobile phase reservoir 5, start the pressure pump in the one-dimensional pumping system 7, switch the one-dimensional multi-position multi-way valve 6 so that the one-dimensional pumping system 7 pumps the tobacco crude extract and the one-dimensional mobile phase into the one-dimensional chromatographic column 8 in sequence and maintains the supply of the one-dimensional mobile phase. At this time, the inlet of the distribution main valve 12 is connected to the waste outlet 24 connected to the waste liquid pool 14, and the one-dimensional chromatographic column 8 is in a state of sample loading and separation and elution and removal of impurities. After the front-stage solvent and impurities are discharged, the one-dimensional chromatographic column 8 begins to elute the nicotine-containing fraction;

[0047] (3) As the one-dimensional pumping system 7 continuously outputs the pure water mobile phase, the nicotine on the one-dimensional chromatographic column 8 is separated and eluted, and the one-dimensional detector 9 indicates a nicotine signal. At this time, the regulating liquid pumping system 11 is immediately turned on, and the regulating liquid pumping system 11 pumps the elution regulating liquid into the mixer 10, so that the elution regulating liquid and the one-dimensional flow fraction containing nicotine are fully mixed in the mixer 10. At the same time, the outlet of the distribution main valve 12 is connected to the first inlet 20 of the multi-position multi-way valve in the left two-dimensional separation channel 2, and the one-dimensional nicotine flow fraction after pH adjustment in the mixer 10 is pumped into the two-dimensional multi-position multi-way valve 13 in the left two-dimensional separation channel 2 through the first inlet 20, and finally flows into the capture column 15 and is captured by the capture column 15. At this time, the left two-dimensional separation channel 2 enters the working state one from the stop state;

[0048] (4) After the one-dimensional nicotine fraction is enriched in the capture column 15 of the left two-dimensional separation channel 2, the communication position of the distribution main valve 12 is switched to connect the first inlet 20 of the right two-dimensional separation channel 3 with the inlet of the distribution main valve 12, and then the first inlet 20 and the first port 22 of the two-dimensional multi-position multi-way valve 13 of the right two-dimensional separation channel 3 are connected. At this time, the one-dimensional nicotine fraction begins to flow into the right two-dimensional separation channel 3, and the one-dimensional nicotine fraction begins to be captured by the capture column 15 in the right two-dimensional separation channel 3; at this time, the right two-dimensional separation channel 3 enters the working state one from the stop state;

[0049] (5) While switching the connection position of the distribution main valve 12 in step (4), the connection position in the left two-dimensional separation channel 2 is switched so that the first port 22 is connected to the chromatographic column port 25, and the second port 23 is connected to the second inlet 21, and the two-dimensional pumping system 16 in the left two-dimensional separation channel 2 is turned on. The two-dimensional pumping system 16 pumps the one-dimensional flow fraction enriched in the capture column 15 and the two-dimensional mobile phase into the two-dimensional chromatographic column 17 to perform two-dimensional separation of the one-dimensional flow fraction. Then, the two-dimensional flow fraction separated by the two-dimensional chromatographic column 17 enters the two-dimensional detector 18 for detection. According to the detection signal of the two-dimensional detector 18, it is selected to connect to the two-dimensional collector 19 for collection or to connect to the waste liquid pool 14 for impurity removal and waste discharge. The two-dimensional nicotine flow fraction can finally enter the two-dimensional collector 19 to complete collection. At this time, the left two-dimensional separation channel 2 enters the working state 2;

[0050] (6) After step (4) is completed and fully captured, the one-dimensional nicotine fraction is enriched in the capture column 15 of the right two-dimensional separation channel 3. At this time, the connection position of the distribution main valve 12 is switched so that the inlet of the distribution main valve 12 is connected to the waste outlet 24. At this time, the one-dimensional chromatography system 1, the left two-dimensional separation channel 2, and the right two-dimensional separation channel 3 are not connected to each other. All three can be operated simultaneously, so steps (7) and (8) can be performed simultaneously.

[0051] (7) Switching the connection position in the right two-dimensional separation channel 3 so that the first port 22 is connected to the chromatographic column port 25, and the second port 23 is connected to the second inlet 21, and the two-dimensional pumping system 16 in the right two-dimensional separation channel 3 is turned on. The two-dimensional pumping system 16 pumps the one-dimensional flow fraction enriched in the capture column 15 and the two-dimensional mobile phase into the two-dimensional chromatographic column 17 to perform two-dimensional separation of the one-dimensional flow fraction. Then, the two-dimensional flow fraction separated by the two-dimensional chromatographic column 17 enters the two-dimensional detector 18 for detection, and finally, according to the detection signal of the two-dimensional detector 18, it is selected to connect to the two-dimensional collector 19 for collection or to connect to the waste liquid pool 14 for impurity removal and waste discharge. The two-dimensional nicotine flow fraction can finally enter the two-dimensional collector 19 to complete collection. At this time, the right two-dimensional separation channel 3 enters the working state II;

[0052] (8) The one-dimensional chromatography system 1 is subjected to post-impurity removal to completely elute and discharge the adsorbed impurities in the one-dimensional chromatography column 8. At this time, the one-dimensional pumping system 7 is turned on, and the pumping system is made to pump the cleaning liquid to clean the one-dimensional chromatography system 1. After the cleaning is completed, the one-dimensional chromatography column 8 can be re-prepared for one-dimensional fractions. The preparation steps of the new one-dimensional fractions are the same as those of steps (1) and (2). During the post-impurity removal cleaning, the inlet of the distribution main valve 12 is continuously connected to the waste outlet 24; in this step, the one-dimensional chromatography system 1 can be left to stand after the cleaning is completed, or the one-dimensional fractions can be re-prepared. During the re-preparation, until the one-dimensional detector 9 detects the nicotine signal, if the left and right two-dimensional separation channels are still in the self-running state of two-dimensional separation, the one-dimensional pumping system 1 and the regulating liquid pumping system 11 need to be shut down to prevent the loss of one-dimensional fractions, wait for the separation and impurity removal of the two-dimensional separation channel to be completed, and then continue to capture a new batch of one-dimensional fractions and prepare two-dimensional fractions;

[0053] (9) While step (8) is being performed, both the left and right two-dimensional separation channels are in the independent self-operation stage of working state 2. Since the self-operation time of the left two-dimensional separation channel 2 is earlier than that of the right two-dimensional separation channel 3, the left two-dimensional separation channel 2 can complete the preparation and collection of the two-dimensional fractions before the right two-dimensional separation channel 3. After the two-dimensional fractions in the left two-dimensional separation channel 2 are prepared, the left two-dimensional separation channel is first cleaned by removing impurities. At this time, the two-dimensional collector 19 is replaced with the waste liquid pool 14. The cleaning liquid is pumped through the two-dimensional pumping system 16 in the left two-dimensional separation channel 2 to clean the channel components, especially the two-dimensional chromatographic column 17, so that the two-dimensional chromatographic column 17 in the left two-dimensional separation channel 2 can be reused. After cleaning, the left two-dimensional separation channel 2 can start to prepare two-dimensional fractions again;

[0054] (10) After the re-preparation of the one-dimensional fraction in step (8) is completed and the impurity removal and cleaning of the left two-dimensional separation channel 2 in step (9) is also completed, the outlet of the distribution main valve 20 is connected to the two-dimensional multi-position multi-way valve 13 in the left two-dimensional separation channel 2, so that the left two-dimensional separation channel 2 can re-enrich and receive the one-dimensional fraction; at this time, the right two-dimensional separation channel 3 is still in the second working state and self-operates or the collection is completed, and the impurity removal and cleaning are started; after the impurity removal and cleaning of the right two-dimensional separation channel 3 are completed and the left two-dimensional separation channel 2 is enriched with the one-dimensional fraction for the second time, the outlet of the distribution main valve 20 is switched to be connected to the right two-dimensional separation channel 3, so that the right two-dimensional separation channel 3 receives the new one-dimensional fraction and enters the working state 2, at this time, the content in step (4) can be repeated, the left two-dimensional separation channel 2 enters the working state 2 and self-operates, and the right two-dimensional separation channel enters the working state 1 to enrich the one-dimensional fraction, and then steps (5) to (10) can be repeated;

[0055] (11) Repeat the above steps (1) to (10) for multiple cycles;

[0056] (12) The two-dimensional fraction containing nicotine collected in the two-dimensional collector 19 is subjected to solid phase extraction to remove the buffer salt, and then concentrated under reduced pressure and freeze-dried to remove methanol and water, thereby obtaining a nicotine freeze-dried powder with a purity of more than 96% and a product yield of 80%.

[0057] According to the above steps, the operation process of the two-dimensional liquid chromatography separation system in the present embodiment under an ideal state is as follows: after the one-dimensional chromatography system 1 produces a one-dimensional fraction, the left two-dimensional separation channel 2 is first connected to the one-dimensional chromatography system 1 to enrich the one-dimensional fraction. After the enrichment of the left two-dimensional separation channel 2 is completed, the right two-dimensional separation channel 3 is switched to enrich the one-dimensional fraction. At this time, the left two-dimensional separation channel 2 is switched to self-operation to start two-dimensional separation. After the right two-dimensional separation channel 3 enriches the one-dimensional fraction, the right two-dimensional separation channel 3 is also switched to self-operation to start two-dimensional separation. At this time, the one-dimensional chromatography system can be operated. System 1 is cleaned after removing impurities so as to re-prepare one-dimensional fractions, and because the two-dimensional separation channels are disconnected, new samples can be directly replaced; then the left two-dimensional separation channel 2 that has completed the two-dimensional separation is cleaned after removing impurities, so that the left two-dimensional separation channel 2 can re-enrich new one-dimensional fractions, and at the same time the right two-dimensional separation channel 3 that has completed the two-dimensional separation is cleaned after removing impurities, and then the left two-dimensional separation channel 2 enriches the new one-dimensional fractions and then the right two-dimensional separation channel 3 is switched to enrich the new one-dimensional fractions, and the steps are repeated continuously and orderly, the process is compact, and time waste is reduced.

[0058] When the one-dimensional mobile phase is replaced with ethanol, and the two-dimensional mobile phase is replaced with: phase A is ethanol, phase B is 0.1% acetic acid aqueous solution, and the two-dimensional elution gradient time and percentage are changed, by repeating the above steps, the solanesol solution can be separated and purified in the tobacco crude extract through the left two-dimensional separation channel 2, and the chlorogenic acid solution can be separated and purified through the right two-dimensional separation channel 3. The separation process is similar to the above process. Specifically, as the one-dimensional chromatographic column 8 is eluted, the one-dimensional detector 9 detects the solanesol signal, and immediately connects the one-dimensional chromatography system 1 with the left two-dimensional separation channel 2 to capture the one-dimensional flow fraction containing solanesol. Then, when the one-dimensional detector 9 detects the chlorogenic acid signal, the one-dimensional chromatography system 1 is immediately connected to the right two-dimensional separation channel 3 to capture the one-dimensional flow fraction containing chlorogenic acid. At the same time, the left two-dimensional separation channel 2 is self-operated to separate and purify solanesol, and then the right two-dimensional separation channel 3 is self-operated to separate and purify chlorogenic acid. The main difference between the two two-dimensional separation channels is that the mobile phases and proportions used are different. Therefore, the two-dimensional liquid chromatography separation system in this embodiment can be applied to the simultaneous separation and preparation of multiple targets in a complex mixed system.

[0059] It can be seen from the above steps that the two two-dimensional separation channels can enter the independent working state 2 after completing the enrichment of the one-dimensional flow fraction in the working state 1, so as to independently and self-operate to perform the two-dimensional separation of the one-dimensional flow fraction, and the two two-dimensional separation channels not only have the working state 1 and the working state 2, but also have a stop state. When the two-dimensional separation channel is in the stop state, the first inlet 20 of the two-dimensional multi-position multi-way valve 13 on the two-dimensional separation channel is disconnected from the outlet of the corresponding distribution main valve 12, and the two-dimensional pumping system 16 on the two-dimensional separation channel at this time can be used to pump the cleaning liquid to clean each channel component in the two-dimensional separation channel, so that the two-dimensional separation channel is free of impurities. The two-dimensional separation channel in the stop state does not interfere with other two-dimensional separation channels and the one-dimensional chromatography system 1, so its internal components can also be disassembled and replaced in the stop state, and the two-dimensional multi-position multi-way valve 13 can also switch the connection position arbitrarily.

[0060] The two-dimensional liquid chromatography separation system in this embodiment can directly replace the added sample and mobile phase in the one-dimensional chromatography system 1 when the left two-dimensional separation channel 2 enters the working state 2, and use the right two-dimensional separation channel 3 to connect with the one-dimensional chromatography system 1. The two two-dimensional separation channels do not affect each other, and can independently and efficiently separate different types and large quantities of samples. The two-dimensional liquid chromatography system is independent, stable, and switches quickly, and has efficient reactions, which improves the processing efficiency. It is suitable for large-scale and multi-type sample separation, and effectively solves the technical problems of low efficiency and poor separation effect of two-dimensional liquid chromatographs in the prior art in separating different types of samples.

[0061] Specific embodiment 2 of the two-dimensional liquid chromatography separation system provided by the present invention:

[0062] The difference from Example 1 is that in Example 2, three two-dimensional separation channels are connected to the distribution main valve, and the distribution main valve is a four-position five-way valve. In other embodiments, the two-dimensional separation channels connected to the distribution main valve can be increased to more than three, so as to be able to separate samples in larger batches, and the number of connection positions and ports of the distribution main valve is increased accordingly.

[0063] Specific embodiment 3 of the two-dimensional liquid chromatography separation system provided by the present invention:

[0064] The difference from Example 1 is that in this Example 3, only one end of the capture column in the two-dimensional separation channel is connected to the two-dimensional multi-position multi-way valve, that is, the capture column has only one port, and the other end of the capture column is connected to the two-dimensional chromatographic column and the two-dimensional pumping system through an external bifurcated pipeline. When the two-dimensional separation channel is in working state two, the capture column is driven by the two-dimensional separation channel through the bifurcated pipeline to supply liquid to the two-dimensional chromatographic column.

[0065] Specific embodiment 4 of the two-dimensional liquid chromatography separation system provided by the present invention:

[0066] The difference from Example 1 is that in Example 4, the collector and the waste liquid pool in the collection pipeline are connected to the rear end of the two-dimensional chromatographic column through a switching valve. In other embodiments, the serial connection mode of the collection pipeline and its internal components can also be changed, as long as the purpose of collecting the two-dimensional separation is met, for example, only a collector is set or an extraction device is directly set to extract the two-dimensional separation to obtain the desired substance.

[0067] Specific embodiment 5 of the two-dimensional liquid chromatography separation system provided by the present invention:

[0068] The difference from Example 1 is that in this Example 5, the one-dimensional multi-position multi-way valve and the one-dimensional pumping system are eliminated in the one-dimensional chromatography system, and the sample and mobile phase are directly added to the one-dimensional chromatography column through an external pipeline. In other embodiments, the components in the one-dimensional chromatography system can also be changed to meet the purpose of being able to supply one-dimensional flow fractions to the distribution main valve, for example, the one-dimensional chromatography system in the prior art can be directly used.

[0069] Specific embodiment 6 of the two-dimensional liquid chromatography separation system provided by the present invention:

[0070] The difference from Example 1 is that in Example 6, in order to reduce costs, part of the separated substances can be extracted for detection, and no detector is set in the one-dimensional chromatography system and each two-dimensional separation channel. Alternatively, in other embodiments, only the two-dimensional separated substances can be detected, and only the two-dimensional detector is set in the two-dimensional separation channel.

[0071] Specific embodiment 7 of the two-dimensional liquid chromatography separation system provided by the present invention:

[0072] The difference from Example 1 is that in this Example 7, the mixer is arranged between the distribution main valve and each two-dimensional multi-position multi-way valve, and the one-dimensional separation products are mixed after being distributed by the distribution main valve, thereby accelerating the one-dimensional separation speed.

[0073] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions recorded in the aforementioned embodiments without creative work, or replace some of the technical features therein with equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present invention shall be included in the protection scope of the present invention.

Claims

1. A two-dimensional liquid chromatography separation system, comprising a one-dimensional chromatography system and a two-dimensional separation channel, wherein a distribution main valve is provided at the tail end of the one-dimensional chromatography system, wherein the distribution main valve is a multi-position multi-way valve, and the distribution main valve comprises an inlet and a plurality of outlets, It is characterized in that There are at least two two-dimensional separation channels, each of which includes a two-dimensional multi-position multi-way valve, a capture column, a two-dimensional pumping system and a two-dimensional chromatographic column for communicating with each outlet of the distribution main valve, the capture column is used to enrich the one-dimensional flow fraction, and the two-dimensional chromatographic column is used to separate the one-dimensional flow fraction enriched by the capture column, the two-dimensional multi-position multi-way valve includes a first inlet, a second inlet and a plurality of ports, the first inlet of the two-dimensional multi-position multi-way valve is connected to the outlet of the distribution main valve, the two-dimensional pumping system is connected to the second inlet, and the capture column and the two-dimensional chromatographic column are correspondingly connected to the corresponding ports of the two-dimensional multi-position multi-way valve; the distribution main valve is connected to the two-dimensional multi-position multi The valves cooperate with each other to realize two working states of the two-dimensional separation channel. Working state one: the inlet of the distribution main valve is connected with the first inlet of the two-dimensional multi-position multi-way valve, and the one-dimensional chromatography system is connected with the capture column through the distribution main valve, the first inlet of the two-dimensional multi-position multi-way valve and the corresponding through port, so as to realize the separate connection between the capture column and the one-dimensional chromatography system; Working state two: the inlet of the distribution main valve is disconnected from the first inlet of the two-dimensional multi-position multi-way valve, and the two-dimensional pumping system is connected with the capture column through the second inlet of the two-dimensional multi-position multi-way valve and the corresponding through port, and the capture column is connected with the two-dimensional chromatography column through the corresponding through port, so as to realize the self-operation of the two-dimensional separation channel.

2. The two-dimensional liquid chromatography separation system according to claim 1, It is characterized in that The multiple ports of the two-dimensional multi-position multi-way valve include a capture column port and a chromatographic column port; there are two capture column ports, including a first port and a second port, the first port is connected to one end of the capture column, the second port is connected to the other end of the capture column, and the chromatographic column port is connected to the two-dimensional chromatographic column; when the two-dimensional separation channel is in working state one, the first port is connected to the first inlet, and the capture column can enrich the one-dimensional fraction; when the two-dimensional separation channel is in working state two, the first port is connected to the chromatographic column port, and the second port is connected to the second inlet, so that the two-dimensional pumping system is connected to the capture column and the two-dimensional chromatographic column, so as to pump the one-dimensional fraction enriched by the capture column into the two-dimensional chromatographic column.

3. The two-dimensional liquid chromatography separation system according to claim 2, It is characterized in that The multiple ports of the two-dimensional multi-position multi-way valve also include a waste discharge port, which is used to discharge waste liquid and impurities. When the two-dimensional separation channel is in working state one, the waste discharge port is connected to the second port.

4. The two-dimensional liquid chromatography separation system according to claim 3, It is characterized in that The waste outlet is connected with a waste liquid pool.

5. The two-dimensional liquid chromatography separation system according to claim 3, It is characterized in that The two-dimensional separation channel also has a stop state. When the two-dimensional separation channel is in the stop state, the first inlet of the two-dimensional multi-position multi-way valve on the two-dimensional separation channel is disconnected from the outlet of the corresponding distribution main valve. At this time, the two-dimensional pumping system on the two-dimensional separation channel is used to pump cleaning liquid to clean each channel component in the two-dimensional separation channel.

6. The two-dimensional liquid chromatography separation system according to any one of claims 1 to 5, It is characterized in that The two-dimensional separation channel also includes a collecting pipeline connected to the two-dimensional chromatographic column; the collecting pipeline includes a collector and a waste liquid pool connected in parallel to the rear end of the two-dimensional chromatographic column.

7. The two-dimensional liquid chromatography separation system according to any one of claims 1 to 5, It is characterized in that The one-dimensional chromatography system comprises a one-dimensional multi-position multi-way valve and a one-dimensional chromatography column. The one-dimensional multi-position multi-way valve comprises a plurality of inlets and an outlet connected to the front end of the one-dimensional chromatography column. The inlets of the one-dimensional multi-position multi-way valve are respectively connected to an injector and a mobile phase reservoir. The one-dimensional multi-position multi-way valve is used to introduce the initial sample and the mobile phase in the injector into the one-dimensional chromatography column when changing positions. The rear end of the one-dimensional chromatography column is connected to the inlet of the distribution main valve.

8. The two-dimensional liquid chromatography separation system according to claim 7, It is characterized in that A one-dimensional pumping system is connected between the front end of the one-dimensional chromatographic column and the outlet of the one-dimensional multi-position multi-way valve.

9. The two-dimensional liquid chromatography separation system according to claim 8, It is characterized in that The one-dimensional chromatography system and each two-dimensional separation channel are equipped with a detector. The one-dimensional detector of the one-dimensional chromatography system is connected between the one-dimensional chromatography column and the distribution main valve, and the two-dimensional detector of each two-dimensional chromatography column is connected to the rear end of the two-dimensional chromatography column.

10. The two-dimensional liquid chromatography separation system according to any one of claims 1 to 5, It is characterized in that A mixer is also arranged between the one-dimensional chromatography system and the inlet of the distribution main valve, and a regulating liquid pumping system is connected to the mixer.

Images