CN112798726A - Analytical circulating multidimensional liquid chromatography separation system - Google Patents

Abstract

The invention provides an analytical circulating multidimensional liquid chromatography separation system which comprises a liquid infusion unit, a sample injection unit, a detection unit, a separation column array, an enrichment column array, a diluent pump, a mixer, a pipeline and a two-position multi-way valve. The invention can effectively reduce the flow path delay volume after the chromatographic separation column, reduce the dissolution, diffusion and mixing of the separated sample, improve the output purity of the sample and contribute to qualitative and quantitative analysis of the complex system sample with high separation difficulty by a liquid chromatography-mass spectrometry system.

Description

Analytical circulating multidimensional liquid chromatography separation system

Technical Field

The invention belongs to the technical field of high performance liquid chromatography separation, and relates to an analytical circulating multidimensional liquid chromatography separation system.

Background

If the sample entering the mass spectrum in the LC-MS is not a monomeric compound, the reliability of the mass spectrometry is very low. Therefore, it is necessary to increase the chromatographic peak capacity of the liquid phase system, to increase the separation capacity, and to increase the purity of the sample entering the mass spectrometry for testing.

Chinese patent application CN110346478A discloses a multidimensional liquid chromatography separation system based on a two-position six-way valve, which can realize cyclic multidimensional liquid chromatography separation. However, in this application the sample must be delivered through a mixer that achieves the mixing effect in a certain volume. When the sample is output, the smaller the flow path delay volume after the chromatographic separation column is, the better the flow path delay volume is, so as to reduce the dissolution, diffusion and mixing of the separated sample. There is no disclosure in this application of how to further reduce the flow path delay volume after the chromatography column array to better improve the purity of the sample output.

Disclosure of Invention

The invention aims to construct an analytical circulating multidimensional liquid chromatography separation system for switching enrichment columns under the real-time guidance of chromatographic signals based on a two-position multi-way valve (the number of ports of the two-position multi-way valve is more than or equal to 8), an enrichment column array and a separation column array. When the sample is output, the sample does not flow through the mixer, so that the flow path delay volume behind the chromatographic separation column can be effectively reduced, the dissolution, diffusion and mixing of the separated sample are reduced, the chromatographic peak broadening behind the separation column is reduced, and the purity of the output sample is improved.

In order to achieve the purpose, the technical scheme of the invention is as follows:

an analytical circulating multidimensional liquid chromatography separation system comprises a liquid infusion unit, a sample introduction unit, a detection unit, a separation column array, an enrichment column array, a diluent pump, a mixer, a pipeline and a two-position multi-way valve, wherein the number of ports of the two-position multi-way valve is more than or equal to 8.

The infusion unit is used for completing the supply of a liquid chromatography separation gradient elution mobile phase; the sample introduction unit is used for introducing samples and is provided with an inlet and an outlet; the detection unit is used for detecting chromatographic signals in the separation process; said array of separation columns having an inlet and an outlet; said enrichment column array having an inlet and an outlet; the two-position multi-way valve has two states of a state A and a state B.

When the analytical circulating multidimensional liquid chromatographic separation system carries out chromatographic separation, when the two-position multi-way valve is in the state A, the mobile phase output by the infusion unit flows into the inlet of the sample injection unit; the solution flowing out of the outlet of the sample injection unit flows into one port of the two-position multi-way valve connected with the outlet of the sample injection unit, passes through the two-position multi-way valve and flows into the inlet of the separation column array connected with one port of the two-position multi-way valve; the solution flowing out of the outlet of the separation column array is detected by the detection unit and flows into one port of the two-position multi-way valve; the detected solution flows out from one port of the two-position multi-way valve through the two-position multi-way valve, is diluted by the diluent output by the diluent pump, is mixed by the mixer and flows into one port of the two-position multi-way valve; diluting the mixed solution, and flowing the diluted mixed solution into an inlet of an enrichment column array connected with one port of a two-position multi-way valve through the two-position multi-way valve; and the solution flowing out of the enrichment column array outlet flows into one port of the two-position multi-way valve connected with the enrichment column array outlet, passes through the two-position multi-way valve and flows out of one port of the two-position multi-way valve.

When the analytical circulating multidimensional liquid chromatographic separation system carries out chromatographic separation, when the two-position multi-way valve is in a state B, the mobile phase output by the infusion unit flows into the inlet of the sample injection unit; the solution flowing out of the outlet of the sample injection unit flows into one port of the two-position multi-way valve connected with the outlet of the sample injection unit, passes through the two-position multi-way valve and flows into the inlet of the enrichment column array connected with one port of the two-position multi-way valve; the solution flowing out of the enrichment column array outlet flows into one port of the two-position multi-way valve connected with the enrichment column array outlet, passes through the two-position multi-way valve and flows into the inlet of the separation column array connected with one port of the two-position multi-way valve; the solution flowing out of the outlet of the separation column array is detected by the detection unit, flows into one port of the two-position multi-way valve, passes through the two-position multi-way valve and flows out of one port of the two-position multi-way valve.

The full online detection circulation multidimensional liquid chromatography separation function is realized by controlling the switching of the state A and the state B of the two-position multi-way valve.

The sample introduction unit can be connected in a connecting pipeline before the inlet of the separation column array or in a bypass of the enrichment column array.

The separation column array is formed by connecting a plurality of chromatographic separation columns in parallel, and only one chromatographic separation column can be conducted at the same time; at least one bypass, the bypass being connected in parallel with the separation column; when the bypass is conducted, the separation column cannot be conducted, and when the separation column is conducted, the bypass cannot be conducted; a fixed inlet and a fixed outlet are arranged on the outside. The separation column array can be constructed by a multi-position selection valve or a two-position four-way valve, and can also be constructed by a two-way valve.

The enrichment column array is formed by connecting a plurality of chromatographic enrichment columns in parallel, and only one enrichment column can be conducted at the same time; at least one bypass, the bypass being connected in parallel with the enrichment column; when the bypass is conducted, the enrichment column cannot be conducted, and when the enrichment column is conducted, the bypass cannot be conducted; two interfaces are arranged outside, namely an interface X and an interface Y, one is an inlet, and the other is an outlet. The enrichment column array can be constructed by a multi-position selection valve or a two-position four-way valve, and can also be constructed by a two-way valve.

The sample introduction unit is a sample introduction device, can be in the form of a sample introduction valve, an automatic sample introduction device or a sample introduction pump and the like, has two states, namely a sample introduction state and a non-sample introduction straight-through state, and is provided with an inlet and an outlet for system flow path connection.

The infusion unit can be composed of a high performance liquid chromatography gradient pump A, a high performance liquid chromatography gradient pump B and a gradient mixer. The diluent pump is a high-efficiency liquid phase diluent pump, and is a unit pump or a multi-element pump. The high performance liquid chromatography gradient pump A, the high performance liquid chromatography gradient pump B and a diluent pump, wherein the diluent can be water, salt solution, methanol, acetonitrile, acetone, ethanol or normal alkane solvent, and the eluent can be methanol, acetonitrile, ethanol, water and mixture thereof, normal alkane and other common organic solvents.

The detection unit is various devices for detecting chromatographic signals in the separation process, including but not limited to an ultraviolet detector, a diode array detector, an evaporative light scattering detector or a mass spectrum detector, and comprises a combined detection system consisting of a plurality of detectors.

The chromatographic columns of the separation column array and the enrichment column array can be selected from the same or different fillers, and the fillers can be silica gel, reversed phase silica gel matrix fillers with C18, Xion, C8, CN groups or amino groups, or various fillers such as macroporous adsorption resin, ion exchange resin and the like.

Compared with the prior art, the invention has the following innovation points and beneficial effects:

when the two-position multi-way valve is in a B state, a detected sample is directly output without a mixer, the flow path delay volume after the separation column array can be effectively reduced, the dissolution, diffusion and mixing of the separated sample are reduced, the chromatographic peak broadening after the separation column is reduced, the output purity of the sample is improved, and the qualitative and quantitative analysis of a complex system sample with high separation difficulty by a liquid chromatography-mass spectrometry system is facilitated; meanwhile, the multi-dimensional liquid chromatography separation system constructed by the invention has the smallest volume of the flow path of the system.

Drawings

FIG. 1 is a flow chart of a system in which a two-position eight-way valve of a multi-dimensional liquid chromatography separation system is in a state A according to a connection mode of the invention (A);

FIG. 2 is a flow chart of a system in which the two-position eight-way valve of the multi-dimensional liquid chromatography separation system is in a B state, the enrichment column is eluted in the positive direction, and the dashed line part is in a non-flow state, according to the connection mode of the invention (A);

FIG. 3 is a flow chart of the system in the state A of the two-position eight-way valve of the multi-dimensional liquid chromatography separation system provided by the connection mode of the invention (B);

FIG. 4 is a flow chart of a system in which the two-position eight-way valve of the multi-dimensional liquid chromatography separation system is in a state B, the enrichment column is in reverse elution, and a dotted line part is in a non-flow state, according to the connection mode of the invention (B);

FIG. 5 is a schematic view of the piping connections of the separation column array;

FIG. 6 is a view showing a structure of a piping connection of the enrichment column array;

FIG. 7(a) is a structural diagram of a multi-dimensional HPLC separation system according to an embodiment of the present invention, wherein the two-position eight-way valve is in the A state;

in fig. 7 (a): 1, an infusion unit (comprising a pump A, a pump B and a mixer A), 2, a sample introduction unit, 3, a separation column array, 4, a detection unit, 5, a diluent pump, 6, a mixer (mixer B), 7, an enrichment column array and 8 two-position eight-way valves;

fig. 7(B) is a structural diagram of a multidimensional high performance liquid chromatography separation system according to an embodiment of the present invention, in which the two-position eight-way valve is in a B state, and a dashed line portion is a pipeline in a non-flow state.

Detailed Description

The following examples are given for the purpose of illustrating the invention only and are not to be construed as limiting the scope of the invention, which is defined in the appended claims.

The implementation mode is as follows:

an analytical circulating multidimensional liquid chromatography separation system comprises a liquid infusion unit, a sample introduction unit, a detection unit, a separation column array, an enrichment column array, a diluent pump, a mixer, a pipeline and a two-position multi-way valve. For simplicity, the two-position multi-way valve is a two-position eight-way valve, the detection unit is an ultraviolet detector, and the sample injection unit is a sample injection valve or an automatic sample injector, which explains the best implementation mode.

A two-position ten-way valve, a two-position twelve-way valve, or a valve group consisting of a plurality of valves, and the like are connected into a two-position eight-way valve, which can be implemented as such; other connection means may be used to achieve the same technical effect.

The 8 ports of the two-position eight-way valve are respectively a port I, a port II, a port III, a port IV, a port V, a.

The pipelines are connected according to any one of the following connection modes (A) to (B):

(A) the outlet of the transfusion unit is connected with the inlet of the sample injection unit, the outlet of the sample injection unit is connected with the port I of the two-position eight-way valve, the port I of the two-position eight-way valve is connected with the inlet of the separation column array, the outlet of the separation column array is connected with the inlet of the detection unit, and the outlet of the detection unit is connected with the port fifthly of the two-position eight-way valve; the port IV of the two-position eight-way valve is connected with the inlet of the mixer, and the outlet of the diluent pump is connected with the inlet of the mixer; the outlet of the mixer is connected with the port III of the two-position eight-way valve; the port II of the two-position eight-way valve is connected with the port X of the enrichment column array, and the port Y of the enrichment column array is connected with the port III of the two-position eight-way valve; and sixthly, outputting the separated sample by the two-position eight-way valve.

(B) The outlet of the transfusion unit is connected with the inlet of the sample injection unit, the outlet of the sample injection unit is connected with the port I of the two-position eight-way valve, the port I of the two-position eight-way valve is connected with the inlet of the separation column array, the outlet of the separation column array is connected with the inlet of the detection unit, and the outlet of the detection unit is connected with the port IV of the two-position eight-way valve; the port of the two-position eight-way valve is connected with the inlet of the mixer, and the outlet of the diluent pump is connected with the inlet of the mixer; the outlet of the mixer is connected with the sixth port of the two-position eight-way valve; the port of the two-position eight-way valve is connected with the interface Y of the enrichment column array, and the interface X of the enrichment column array is connected with the port II of the two-position eight-way valve; and the port III of the two-position eight-way valve outputs the separated sample.

When the detection unit is a mass spectrum detector or an evaporative light scattering detector or the like, or when the detector is a detector with flow limitation, the detection unit can be connected into a system flow path through a shunt branch, an active shunt or a passive shunt, or the like. The shunting branch is provided with three interfaces, one of which is an inlet and is connected with the outlet of the separation column array; one is a first outlet (detection outlet from which a small micro-fluid is detected to flow out as a detection window or detection access point) connected to the inlet of the detector; one is the second outlet (from which the main flow exits). In the connection mode (A), the second outlet of the shunt branch is connected with the port fifthly of the two-position eight-way valve; in the connection mode (B), the second outlet of the shunt branch is connected with the port (r) of the two-position eight-way valve.

The two position eight valve of fig. 1 is in state a. The outlet of the transfusion unit is connected with the inlet of the sample injection unit, the outlet of the sample injection unit is connected with the port I of the two-position eight-way valve, the port I of the two-position eight-way valve is communicated with the port I, the port I of the two-position eight-way valve is connected with the inlet of the separation column array, the outlet of the separation column array is connected with the inlet of the detection unit, and the outlet of the detection unit is connected with the port III of the two-position eight-way valve; the port fifthly of the two-position eight-way valve is communicated with the port fourthly, the port fourthly of the two-position eight-way valve is connected with the inlet of the mixer, and the outlet of the diluent pump is connected with the inlet of the mixer; the outlet of the mixer is connected with the port III of the two-position eight-way valve; the third port of the two-position eight-way valve is communicated with the fourth port, the fourth port of the two-position eight-way valve is connected with the interface X of the enrichment column array, and the interface Y of the enrichment column array is connected with the seventh port of the two-position eight-way valve; and the port of the two-position eight-way valve is communicated with the port of the sixth valve, and the separated samples are output by the port of the two-position eight-way valve.

As shown in fig. 1, when the analytical circulating multidimensional liquid chromatography system performs chromatography separation, when the two-position eight-way valve is in the state a, the mobile phase output by the infusion unit flows into the inlet of the sample injection unit; the solution flowing out of the outlet of the sample injection unit flows into a port I of the two-position multi-way valve connected with the outlet of the sample injection unit, flows out of a port I communicated with the port I of the two-position eight-way valve through the two-position multi-way valve, and flows into an inlet of a separation column array connected with the port I of the two-position eight-way valve; the solution flowing out of the outlet of the separation column array is detected by the detection unit and flows into the port fifthly of the two-position eight-way valve; the detected solution flows out from a port IV communicated with a port V in the two-position eight-way valve through the two-position eight-way valve, is diluted by the diluent output by the diluent pump, is mixed by the mixer and flows into a port III of the two-position eight-way valve; diluting the mixed solution, flowing out from a port II communicated with a port III in the two-position eight-way valve through the two-position eight-way valve, and flowing into an inlet (interface X) of an enrichment column array connected with the port II of the two-position eight-way valve; the solution flowing out of the enrichment column array outlet (interface Y) flows into a port (c) in the two-position eight-way valve, which is connected with the enrichment column array outlet, and flows out of a port (c) in the two-position eight-way valve, which is communicated with the port (c).

The two-position eight-way valve in fig. 2 is in the B state. The outlet of the infusion unit is connected with the inlet of the sample injection unit, the outlet of the sample injection unit is connected with the port I of the two-position eight-way valve, the port I of the two-position eight-way valve is communicated with the port II, the port II of the two-position eight-way valve is connected with the interface X of the enrichment column array, and the interface Y of the enrichment column array is connected with the port III of the two-position eight-way valve; the port of the two-position eight-way valve is communicated with the port of the eight-way valve, the port of the eight-way valve of the two-position eight-way valve is connected with the inlet of the separation column array, the outlet of the separation column array is connected with the inlet of the detection unit, and the outlet of the detection unit is connected with the port of the eight-way valve of the two-position eight-way valve; and the port fifthly of the two-position eight-way valve is communicated with the port sixthly, and the separated sample is output from the port sixthly of the two-position eight-way valve.

As shown in fig. 2, when the analytical circulating multidimensional liquid chromatography system performs chromatography separation, when the two-position eight-way valve is in the state B, the mobile phase output by the infusion unit flows into the inlet of the sample injection unit; the solution flowing out of the outlet of the sample injection unit flows into a port I of the two-position multi-way valve connected with the outlet of the sample injection unit, flows out of a port II communicated with the port I of the two-position eight-way valve through the two-position eight-way valve, and flows into an inlet (interface X) of the enrichment column array connected with the port II of the two-position eight-way valve; the solution flowing out of an outlet (interface Y) of the enrichment column array flows into a seventh port connected with the outlet of the enrichment column array in the two-position eight-way valve, flows out of a seventh port communicated with the seventh port in the two-position eight-way valve through the two-position eight-way valve and flows into an inlet of a separation column array connected with the seventh port of the two-position eight-way valve; and the solution flowing out of the outlet of the separation column array is detected by the detection unit, flows into the port fifthly of the two-position eight-way valve, passes through the two-position eight-way valve, and flows out of the port communicated with the port fifthly in the two-position eight-way valve.

The two position eight valve of fig. 3 is in state a. The outlet of the transfusion unit is connected with the inlet of the sample injection unit, the outlet of the sample injection unit is connected with the port I of the two-position eight-way valve, the port I of the two-position eight-way valve is communicated with the port I, the port I of the two-position eight-way valve is connected with the inlet of the separation column array, the outlet of the separation column array is connected with the inlet of the detection unit, and the outlet of the detection unit is connected with the port IV of the two-position eight-way valve; the port IV of the two-position eight-way valve is communicated with the port V of the two-position eight-way valve, the port V of the two-position eight-way valve is connected with the inlet of the mixer, and the outlet of the diluent pump is connected with the inlet of the mixer; the outlet of the mixer is connected with the sixth port of the two-position eight-way valve; the port of the two-position eight-way valve is communicated with the port of the enrichment column array, the port of the two-position eight-way valve is connected with the port Y of the enrichment column array, and the port X of the enrichment column array is connected with the port II of the two-position eight-way valve; and the port II of the two-position eight-way valve is communicated with the port III of the two-position eight-way valve, and the port III of the two-position eight-way valve outputs the separated sample.

As shown in fig. 3, when the analytical circulating multidimensional liquid chromatography system performs chromatography separation, when the two-position eight-way valve is in the state a, the mobile phase output by the infusion unit flows into the inlet of the sample injection unit; the solution flowing out of the outlet of the sample injection unit flows into a port I of the two-position eight-way valve connected with the outlet of the sample injection unit, flows out of a port I of the two-position eight-way valve communicated with the port I through the two-position multi-way valve, and flows into an inlet of a separation column array connected with the port I of the two-position eight-way valve; the solution flowing out of the outlet of the separation column array is detected by the detection unit and flows into the port IV of the two-position eight-way valve; the detected solution flows out from a port fifthly communicated with a port fourteen in the two-position eight-way valve through the two-position eight-way valve, is diluted by the diluent output by the diluent pump, is mixed by the mixer and flows into a port sixthly of the two-position eight-way valve; the diluted and mixed solution flows out of a port seventhly communicated with the port sixthly in the two-position eight-way valve through a two-position eight-way valve and flows into an inlet (interface Y) of an enrichment column array connected with the port seventhly of the two-position eight-way valve; the solution flowing out of the enrichment column array outlet (interface X) flows into a port II of the two-position eight-way valve connected with the enrichment column array outlet, and flows out of a port III of the two-position eight-way valve communicated with the port II through the two-position eight-way valve.

In fig. 4, the two-position eight-way valve is in the B state. The outlet of the infusion unit is connected with the inlet of the sample injection unit, the outlet of the sample injection unit is connected with the port I of the two-position eight-way valve, the port I of the two-position eight-way valve is communicated with the port II, the port II of the two-position eight-way valve is connected with the interface X of the enrichment column array, and the interface Y of the enrichment column array is connected with the port III of the two-position eight-way valve; the port of the two-position eight-way valve is communicated with the port of the eighty valve, the port of the eighty valve of the two-position eight-way valve is connected with the inlet of the separation column array, the outlet of the separation column array is connected with the inlet of the detection unit, and the outlet of the detection unit is connected with the port of the eighty valve of the two-position eight-way valve; and the port IV of the two-position eight-way valve is communicated with the port III of the two-position eight-way valve, and the port III of the two-position eight-way valve outputs the separated sample.

As shown in fig. 4, when the analytical circulating multidimensional liquid chromatography system performs chromatography separation, when the two-position eight-way valve is in the state B, the mobile phase output by the infusion unit flows into the inlet of the sample injection unit; the solution flowing out of the outlet of the sample injection unit flows into a port I of the two-position multi-way valve connected with the outlet of the sample injection unit, flows out of a port II communicated with the port I of the two-position eight-way valve through the two-position eight-way valve, and flows into an inlet (interface X) of the enrichment column array connected with the port II of the two-position eight-way valve; the solution flowing out of an outlet (interface Y) of the enrichment column array flows into a seventh port connected with the outlet of the enrichment column array in the two-position eight-way valve, flows out of a seventh port communicated with the seventh port in the two-position eight-way valve through the two-position eight-way valve and flows into an inlet of a separation column array connected with the seventh port of the two-position eight-way valve; the solution flowing out of the outlet of the separation column array is detected by the detection unit, flows into the port IV of the two-position eight-way valve, passes through the two-position multi-way valve, and flows out of the port III communicated with the port IV from the two-position eight-way valve.

Example (b): production type circulating multidimensional liquid chromatography separation system structure

In the embodiment, a system flow path adopts a connection mode (A), a sample introduction unit is a sample introduction valve, a detection unit is an ultraviolet detector, an enrichment column array is composed of a secondary enrichment column array and operates according to the principle of one enrichment column array, each stage of enrichment column array is provided with 9 enrichment columns, namely, the enrichment column array is 18 enrichment columns which are sequentially numbered as a 1 st enrichment column, a 2 nd enrichment column and the like of the enrichment column array, and the last enrichment column is numbered as an 8 th enrichment column of the enrichment column array; the separation column array comprises 5 separation columns which are sequentially numbered as a 1 st separation column, a 2 nd separation column and the like, and the last separation column is a 5 th separation column; the two-position eight-way valve in fig. 7(a) is in the a state, and the two-position eight-way valve in fig. 7(B) is in the B state.

The following is the three-dimensional separation process control of the above-mentioned multidimensional liquid chromatography separation system structure:

firstly, cleaning an enrichment column and a separation column; and sequentially switching each enrichment column and each separation column into the flow path, and observing signals of the detector to judge the cleaning effect.

Controlling the first-dimension separation process: the two-position eight-way valve is in the state A, and is shown in figure 1; loading a sample into a dosing ring on a sample injection valve; selecting a first dimension chromatographic separation column, e.g., the 1 st separation column, which is manually turned on; when the injection valve is switched to an INJECT state, starting first-dimension separation; under the assistance of a diluent pump, sequentially enriching target fractions by using enrichment columns from 1 st to 9 th of an enrichment column array according to sample properties and detection signals after the first-dimensional liquid phase separation, and reserving enrichment columns from 10 th to 18 th of the enrichment column array for use in third-dimensional separation; repeating the steps until enough compounds exist in the 1 st to the 9 th enrichment columns of the enrichment column array, and switching to the control of the second-dimension separation process.

And (3) controlling a second-dimension loading process: after the control of the first dimension separation process is finished, the sample injection unit should be switched to a non-sample injection straight-through state, the two-position eight-way valve is switched to a B state, and a second dimension chromatographic separation column, for example, the 2 nd separation column, is selected and is manually conducted, see fig. 2; and selecting one enrichment column from the 1 st enrichment column to the 9 th enrichment column of the enrichment column array as a sample column for second-dimensional separation, eluting a target sample in the enrichment column into the 2 nd separation column, and finishing the sample loading process of the second-dimensional separation.

And (3) controlling a second-dimension separation process: after the second-dimension sample loading process is completed, when the two-position eight-way valve is switched to the state A, the second-dimension separation is started, and the reference is made to the graph 1; in the second dimension separation process, under the assistance of a diluent pump, target fractions are sequentially switched to enrichment columns from the 10 th to the 18 th of the enrichment column array for enrichment according to sample properties and detection signals.

And (3) controlling a third-dimensional separation process: after the control of the second dimension separation process is finished, the two-position eight-way valve is switched to the state B, which is shown in figure 2; the sample introduction unit keeps a non-sample introduction straight-through state; selecting a third dimension chromatographic separation column, e.g., the 3 rd separation column, which is manually turned on; selecting one enrichment column of the 10 th to 18 th enrichment columns of the enrichment column array as a sample column for a third-dimensional separation; when the enrichment column is conducted, a third-dimensional separation process is started; the separated sample is directly output without a mixer and an enrichment column array; and repeating the steps to finish the third-dimensional separation of all samples.

Claims (2)

1. An analytical circulating multidimensional liquid chromatography separation system is characterized by comprising a liquid infusion unit, a sample introduction unit, a detection unit, a separation column array, an enrichment column array, a diluent pump, a mixer, a pipeline and a two-position multi-way valve; the number of the ports of the two-position multi-way valve is more than or equal to 8;

the infusion unit is used for completing the supply of a liquid chromatography separation gradient elution mobile phase;

the sample introduction unit is used for introducing samples and is provided with an inlet and an outlet;

the detection unit is used for detecting chromatographic signals in the separation process;

said array of separation columns having an inlet and an outlet;

said enrichment column array having an inlet and an outlet;

the two-position multi-way valve has two states, namely a state A and a state B;

when the analytical circulating multidimensional liquid chromatographic separation system carries out chromatographic separation, when the two-position multi-way valve is in the state A, the mobile phase output by the infusion unit flows into the inlet of the sample injection unit; the solution flowing out of the outlet of the sample injection unit flows into one port of the two-position multi-way valve connected with the outlet of the sample injection unit, passes through the two-position multi-way valve and flows into the inlet of the separation column array connected with one port of the two-position multi-way valve; the solution flowing out of the outlet of the separation column array is detected by the detection unit and flows into one port of the two-position multi-way valve; the detected solution flows out from one port of the two-position multi-way valve through the two-position multi-way valve, is diluted by the diluent output by the diluent pump, is mixed by the mixer and flows into one port of the two-position multi-way valve; diluting the mixed solution, and flowing the diluted mixed solution into an inlet of an enrichment column array connected with one port of a two-position multi-way valve through the two-position multi-way valve; the solution flowing out of the enrichment column array outlet flows into one port of the two-position multi-way valve connected with the enrichment column array outlet, passes through the two-position multi-way valve and flows out of one port of the two-position multi-way valve;

when the analytical circulating multidimensional liquid chromatographic separation system carries out chromatographic separation, when the two-position multi-way valve is in a state B, the mobile phase output by the infusion unit flows into the inlet of the sample injection unit; the solution flowing out of the outlet of the sample injection unit flows into one port of the two-position multi-way valve connected with the outlet of the sample injection unit, passes through the two-position multi-way valve and flows into the inlet of the enrichment column array connected with one port of the two-position multi-way valve; the solution flowing out of the enrichment column array outlet flows into one port of the two-position multi-way valve connected with the enrichment column array outlet, passes through the two-position multi-way valve and flows into the inlet of the separation column array connected with one port of the two-position multi-way valve; the solution flowing out of the outlet of the separation column array is detected by the detection unit, flows into one port of the two-position multi-way valve, passes through the two-position multi-way valve and flows out of one port of the two-position multi-way valve;

the full online detection circulation multidimensional liquid chromatography separation function is realized by controlling the switching of the state A and the state B of the two-position multi-way valve.

2. An analytical circulating multidimensional liquid chromatography separation system as claimed in claim 1 wherein the sample introduction unit is connected in a connecting line before the inlet of the separation column array or in a bypass of the enrichment column array.

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