CN215415249U

CN215415249U - Production type circulating multidimensional liquid chromatography separation system

Info

Publication number: CN215415249U
Application number: CN202022669058.9U
Authority: CN
Inventors: 李宜珊
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-11-10
Filing date: 2020-11-10
Publication date: 2022-01-04
Anticipated expiration: 2030-11-10

Abstract

The utility model provides a production type circulating multidimensional liquid chromatography separation system which comprises a liquid conveying unit, a sample injection unit, a detection unit, a separation column array, an enrichment column array, a two-position four-way valve, a diluent pump, a mixer, 4 two-way valves, 4 three-way valves and connecting pipelines. Based on the same infusion unit and the same detection unit, the diluent pump is used for assisting the enrichment or the trapping of the compound, and the full online monitoring and control of the multi-dimensional chromatographic separation can be realized. The utility model realizes the high-efficiency separation of the monomer compounds in the complex system sample with high separation difficulty by selecting different chromatographic stationary phases and mobile phases.

Description

Production type circulating multidimensional liquid chromatography separation system

Technical Field

The utility model belongs to the technical field of high performance liquid chromatography separation, and relates to a production type circulating multidimensional liquid chromatography separation system.

Background

In order to perform automatic, large-scale and systematic separation and purification on complex samples with uneven content and unknown components and quickly perform high-resolution high-throughput qualitative and quantitative analysis on all the components, it is necessary to establish a multi-dimensional chromatographic separation system and a multi-dimensional separation method. In addition, in order to realize high-resolution separation of substances which are similar in physicochemical properties and difficult to separate, such as isomers and structural analogs, it is also necessary to establish a cyclic multidimensional separation system and a separation method.

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. When preparing and collecting separated sample components, the smaller the flow path delay volume after the chromatographic separation column is, the better the flow path delay volume is, and the dissolution and mixing of the separated samples are avoided. However, this application does not disclose how to further reduce the flow path delay volume after the chromatography column array to better improve the purity and yield of sample collection.

SUMMERY OF THE UTILITY MODEL

The utility model aims to construct a production type circulating multidimensional liquid chromatography separation system for switching enrichment columns under the real-time guidance of chromatographic signals based on a two-position four-way valve, an enrichment column array and a separation column array, so that the flow path delay volume behind the chromatographic separation columns can be effectively reduced when separated sample components need to be collected, and the purity and yield of sample collection are improved.

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

a production type circulating multidimensional liquid chromatography separation system comprises a transfusion unit, a sample introduction unit, a detection unit, a separation column array, an enrichment column array, a two-position four-way valve, a diluent pump and a mixer; the two-position four-way valve is provided with 4 interfaces including a first position, a second position, a third position and a fourth position; the detection unit is used for detecting chromatographic signals in the separation process; the sample introduction unit is used for sample introduction; the liquid phase gradient pump unit is used for completing the supply of the chromatographic separation gradient elution mobile phase.

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 provided for the outside, namely an interface X and an interface Y. 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 production type circulating multidimensional liquid chromatography separation system also comprises two-way valves V1-V4, three-way valves T1-T4 and connecting pipelines.

The connecting pipeline is any one of the following connecting modes (A) to (C):

(A) 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 position I of the two-position four-way valve, the position II of the two-position four-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 a first interface of the tee T1; a second interface of the tee T1 is connected with one end of a two-way valve V1, and the other end of the two-way valve V1 is output to the waste liquid barrel; a third interface of the tee T1 is connected with a first interface of the tee T2, a second interface of the tee T2 is connected with an outlet of the diluent pump, a third interface of the tee T2 is connected with an inlet of the mixer, and an outlet of the mixer is connected with one end of a two-way valve V2; the other end of the two-way valve V2 is connected with a first interface of a tee T3, a second interface of the tee T3 is connected with one end of a two-way valve V3, and the other end of the two-way valve V3 is connected with the third position of the two-position four-way valve; a third interface of the tee T3 is connected with an interface Y of the enrichment column array, and an interface X of the enrichment column array is connected with a first interface of the tee T4; the second interface of the tee T4 is connected with the second interface of the two-position four-way valve, the third interface of the tee T4 is connected with one end of the two-way valve V4, and the other end of the two-way valve V4 is output to the waste liquid barrel.

(B) 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 position I of the two-position four-way valve, the position II of the two-position four-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 a first interface of the tee T1; a second interface of the tee T1 is connected with one end of a two-way valve V1, and the other end of the two-way valve V1 is output to the waste liquid barrel; a third interface of the tee T1 is connected with a first interface of the tee T2, a second interface of the tee T2 is connected with an outlet of the diluent pump, a third interface of the tee T2 is connected with an inlet of the mixer, and an outlet of the mixer is connected with one end of a two-way valve V2; the other end of the two-way valve V2 is connected with a first interface of a tee T3, a second interface of the tee T3 is connected with an interface Y of an enrichment column array, and an interface X of the enrichment column array is connected with the second position of the two-position four-way valve; a third interface of the tee T3 is connected with one end of a two-way valve V3, the other end of the two-way valve V3 is connected with a first interface of the tee T4, and a second interface of the tee T4 is connected with a third interface of the two-position four-way valve; and a third interface of the tee T4 is connected with one end of a two-way valve V4, and the other end of the two-way valve V4 is output to the waste liquid barrel.

(C) 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 position I of the two-position four-way valve, the position II of the two-position four-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 a first interface of the tee T1; a second interface of the tee T1 is connected with one end of a two-way valve V1, and the other end of the two-way valve V1 is output to the waste liquid barrel; a third interface of the tee T1 is connected with a first interface of the tee T2, a second interface of the tee T2 is connected with an outlet of the diluent pump, a third interface of the tee T2 is connected with an inlet of the mixer, and an outlet of the mixer is connected with one end of a two-way valve V2; the other end of the two-way valve V2 is connected with a first interface of a tee T3, and a second interface of the tee T3 is connected with a third interface of the two-position four-way valve; a third interface of the tee T3 is connected with one end of a two-way valve V3, and the other end of the two-way valve V3 is connected with a first interface of the tee T4; a second interface of the tee T4 is connected with an interface Y of the enrichment column array, and an interface X of the enrichment column array is connected with the second position of the two-position four-way valve; and a third interface of the tee T4 is connected with one end of a two-way valve V4, and the other end of the two-way valve V4 is output to the waste liquid barrel.

In the connection modes of (A) to (C), the sample introduction unit can also be connected between the inlet of the separation column array and the position (r) of the two-position four-way valve, or in a connecting pipeline of the enrichment column array and the two-position four-way valve, or in a bypass of the enrichment column array.

(A) And (B) the enrichment and elution directions of the enrichment column in the system in the connection mode are opposite, which is called reverse elution, so that certain mobile phase and separation time can be saved.

(C) The enrichment and elution directions of the enrichment column in the connection mode system are consistent, and the system is called forward elution.

Based on the pipeline connection mode of the production type circulating multidimensional liquid chromatography separation system, the system can realize sample loading, separation and enrichment by controlling the state of the two-position four-way valve and the on-off state of the two-way valves V1-V4, and the circulating multidimensional chromatography separation function of full online detection is completed.

When collection is required, the output (waste position) of both two-way valves V1 and V4 can be connected to the collector. The output end of the two-way valve V1 is connected with the collector, so that the flow path delay volume after the chromatographic separation column can be effectively reduced, and the high-purity collection of the separated sample is convenient to realize; the output end of the two-way valve V4 is connected with a collector, and can be used for checking the sample enrichment effect and collecting the sample in the enrichment column. The collector includes a valve-switched collection system and an X-Y matrix collection system.

In the development and optimization stage of the separation method, the output end of the two-way valve V4 can be connected with a temporary detection unit to detect the enrichment effect of the enrichment column in real time.

In the flow path of the circulating multidimensional liquid chromatography separation system, the two-way valve V1 and the three-way valve T1 can be replaced by a two-position three-way valve, the two-way valves V2 and V3 and the three-way valve T3 can be replaced by a two-position three-way valve, and the two-way valve V4 and the three-way valve T4 can be replaced by a two-position three-way valve. The two-position four-way valve can also be constructed by a two-way valve or a two-position three-way valve. The two-way mixer and the three-way T2 can be replaced with a three-way mixer.

The two-way valve has two states of connection and disconnection.

The sample introduction unit is a sample introduction device, can be in the form of a sample introduction valve 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 utility model has the following innovation points and beneficial effects:

the multi-dimensional liquid chromatography separation system is constructed based on a two-position four-way valve, an enrichment column array and a separation column array, the two-way valve is connected behind the detection unit through a tee joint, the flow path delay volume behind the separation column array can be effectively reduced when separated sample components need to be collected, and the purity and yield of sample collection are improved. If a flow path with a large drift diameter is needed, the two-position four-way valve can be constructed by adopting two-way valves, the number of the two-way valves needed by the whole system is 8, and the number of the two-way valves is small.

Drawings

FIG. 1 is a schematic diagram of a pipeline connection structure of a two-position four-way valve of a reverse elution multi-dimensional liquid chromatography separation system in the state A according to the connection mode of the utility model (A);

FIG. 2 is a flow chart of a system in which a two-position four-way valve of a reverse-elution multi-dimensional liquid chromatography separation system in the connection mode of the utility model (A) is in a state A, and a dotted line part is in a non-conduction state;

FIG. 3 is a flow chart of a system in which a two-position four-way valve of a reverse-elution multi-dimensional liquid chromatography separation system in the connection mode (A) of the present invention is in a B state, and a dotted line part is in a non-conduction state;

FIG. 4 is a schematic diagram of the pipeline connection structure of the two-position four-way valve of the reverse elution multi-dimensional liquid chromatography separation system in the A state according to the connection mode of the utility model (B);

FIG. 5 is a flow chart of a system in which a two-position four-way valve of a reverse-elution multi-dimensional liquid chromatography separation system in the connection mode of the utility model (B) is in a state A, and a dotted line part is in a non-conduction state;

FIG. 6 is a flow chart of a system in which a two-position four-way valve of a reverse-elution multi-dimensional liquid chromatography separation system in the connection mode of the utility model (B) is in a B state, and a dotted line part is in a non-conduction state;

FIG. 7 is a schematic diagram of the connection of the two-position four-way valve of the forward elution multi-dimensional liquid chromatography separation system in the A state according to the connection mode of the present invention (C);

FIG. 8 is a flow chart of a system in which a two-position four-way valve of a forward elution multi-dimensional liquid chromatography separation system is in a state A according to the connection mode of the present invention (C), and a broken line portion is in a non-conducting state;

FIG. 9 is a flow chart of a system in which a two-position four-way valve of a forward elution multi-dimensional liquid chromatography separation system is in a B state according to the connection mode of the present invention (C), and a dotted line portion is in a non-conducting state;

FIG. 10 is a schematic diagram of the pipeline connection of the separation column array, in which both ends of each separation column are connected to the common passage through two-way valves, the two-way valves at both ends of each separation column control the connection and disconnection of each separation column, and the two-way valves control the connection and disconnection of the bypass;

fig. 11 is a view showing a piping connection structure of an enrichment column array, in which both ends of each enrichment column are connected to a common passage through two-way valves, each enrichment column is controlled to be turned on and off by the two-way valves at both ends thereof, and a bypass is controlled to be turned on and off by one two-way valve;

FIG. 12(a) is a structural diagram of a multidimensional high performance liquid chromatography separation system according to an embodiment of the present invention, wherein a two-position four-way valve is in an A state;

in fig. 12 (a): 1 transfusion unit (comprising a pump A, a pump B and a mixer 1), 2 sample introduction unit, 3 separation column array, 4 detection unit, 5 diluent pump, 6 mixer (mixer 2), 7 enrichment column array and 8 two-position four-way valve;

fig. 12(B) is a structural diagram of a multidimensional high performance liquid chromatography separation system according to an embodiment of the present invention, and the two-position four-way valve is in a B state.

Detailed Description

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

A production type circulating multidimensional liquid chromatography separation system comprises a transfusion unit, a sample introduction unit, a detection unit, a separation column array, an enrichment column array, a two-position four-way valve, a diluent pump and a mixer; wherein, the enrichment column array and the separation column array are constructed by a multi-position selection valve, and the diluent pump is a high-efficiency liquid phase diluent pump.

In addition, the production type circulating multidimensional liquid chromatography separation system also comprises two-way valves V1-V4, three-way valves T1-T4 and connecting pipelines. The connecting pipeline is any one of the following connecting modes (A) to (C):

In fig. 2, the two-position four-way valve is in the state a, in which the two-way valves V1 and V3 are in the non-conducting state and the two-way valves V2 and V4 are in the conducting state. The outlet of the transfusion unit is connected with the inlet of the sample introduction unit; the sample injection unit can be in a sample injection state or a non-sample injection state according to requirements; the outlet of the sample introduction unit is connected with the first position of the two-position four-way valve; the first position and the fourth position of the two-position four-way valve are communicated and connected with the inlet of the separation column array, and any chromatographic column in the separation column array is selected for separation; the outlet of the separation column array is connected with the inlet of the detection unit, the detection unit detects chromatographic signals, and the outlet of the detection unit is connected with the inlet of the mixer through a tee T1 and a tee T2; the diluent pump is connected with the inlet of the mixer through a tee T2, the mixer dilutes the column and then flows out of the sample, and the outlet of the mixer is connected with one end of a two-way valve V2; the two-way valve V2 is switched on, the two-way valve V2 is connected with an interface Y of the enrichment column array (at the moment, the interface Y of the enrichment column array is the inlet of the enrichment column array) through a tee T3, the enrichment column is selected to enrich the target sample, and an interface X of the enrichment column array (at the moment, the interface X of the enrichment column array is the outlet of the enrichment column array) is connected with the two-way valve V4 through a tee T4; the two-way valve V4 is conducted, and the other end of the two-way valve V4 is output to the waste liquid barrel.

In fig. 3, the two-position four-way valve is in the B state, in which the two-way valves V1 and V3 are in the on state, and the two-way valves V2 and V4 are in the off state. The outlet of the transfusion unit is connected with the inlet of the sample injection unit, and the sample injection unit is in a non-sample injection straight-through state; the outlet of the sample introduction unit is connected with the first position of the two-position four-way valve; the first position and the second position of the two-position four-way valve are communicated and are connected with the interface X of the enrichment column array through a tee T4 (at the moment, the interface X of the enrichment column array is the inlet of the enrichment column array); eluting the target sample in the enrichment column, and finishing loading the target sample to a separation column or separating by using the separation column; an interface Y of the enrichment column array (at the moment, the interface Y of the enrichment column array is an outlet of the enrichment column array) is connected with one end of a two-way valve V3 through a tee T3; the two-way valve V3 is conducted, the other end of the two-way valve V3 is connected with the third position of the two-position four-way valve, and the third position of the two-position four-way valve is conducted with the fourth position; the fourth position of the two-position four-way valve is connected with the inlet of the separation column array; selecting any chromatographic column in the separation column array for separation; the outlet of the separation column array is connected with the inlet of the detection unit, the detection unit detects chromatographic signals, the outlet of the detection unit is connected with one end of a two-way valve V1 through a tee T1, and the other end of the two-way valve V1 is connected with a waste liquid barrel or is connected with the inlet of a collector, so that sample collection is realized.

In fig. 5, the two-position four-way valve is in the state a, in which the two-way valves V1 and V3 are in the non-conducting state and the two-way valves V2 and V4 are in the conducting state. The outlet of the transfusion unit is connected with the inlet of the sample introduction unit; the sample injection unit can be in a sample injection state or a non-sample injection state according to requirements; the outlet of the sample introduction unit is connected with the first position of the two-position four-way valve; the first position and the fourth position of the two-position four-way valve are communicated and connected with the inlet of the separation column array, and any chromatographic column in the separation column array is selected for separation; the outlet of the separation column array is connected with the inlet of the detection unit, the detection unit detects chromatographic signals, and the outlet of the detection unit is connected with the inlet of the mixer through a tee T1 and a tee T2; the diluent pump is connected with the inlet of the mixer through a tee T2, the mixer dilutes the column and then flows out of the sample, and the outlet of the mixer is connected with one end of a two-way valve V2; the two-way valve V2 is conducted, the two-way valve V2 is connected with an interface Y of the enrichment column array (at the moment, the interface Y of the enrichment column array is the inlet of the enrichment column array) through a three-way valve T3, the enrichment column is selected to enrich the target sample, and an interface X of the enrichment column array (at the moment, the interface X of the enrichment column array is the outlet of the enrichment column array) is connected with position two of the two-position four-way valve; the second position of the two-position four-way valve is communicated with the third position, and the third position of the two-position four-way valve is connected with one end of a two-way valve V4 through a tee T4; the two-way valve V4 is conducted, and the other end of the two-way valve V4 is output to the waste liquid barrel.

In fig. 6, the two-position four-way valve is in the B state, in which the two-way valves V1 and V3 are in the on state, and the two-way valves V2 and V4 are in the off state. The outlet of the transfusion unit is connected with the inlet of the sample injection unit, and the sample injection unit is in a non-sample injection straight-through state; the outlet of the sample introduction unit is connected with the first position of the two-position four-way valve; the first position and the second position of the two-position four-way valve are communicated and are connected with the interface X of the enrichment column array through a tee T4 (at the moment, the interface X of the enrichment column array is the inlet of the enrichment column array); eluting the target sample in the enrichment column, and finishing loading the target sample to a separation column or separating by using the separation column; an interface Y of the enrichment column array (at the moment, the interface Y of the enrichment column array is an outlet of the enrichment column array) is connected with one end of a two-way valve V3 through a tee T3; the two-way valve V3 is conducted, the other end of the two-way valve V3 is connected with the third position of the two-position four-way valve through T4, and the third position of the two-position four-way valve is conducted with the fourth position; the fourth position of the two-position four-way valve is connected with the inlet of the separation column array; selecting any chromatographic column in the separation column array for separation; the outlet of the separation column array is connected with the inlet of the detection unit, the detection unit detects chromatographic signals, the outlet of the detection unit is connected with one end of a two-way valve V1 through a tee T1, and the other end of the two-way valve V1 is connected with a waste liquid barrel or is connected with the inlet of a collector, so that sample collection is realized.

In fig. 8, the two-position four-way valve is in the state a, in which the two-way valves V1 and V3 are in the non-conductive state and the two-way valves V2 and V4 are in the conductive state. The outlet of the transfusion unit is connected with the inlet of the sample introduction unit; the sample injection unit can be in a sample injection state or a non-sample injection straight-through state according to requirements; the outlet of the sample introduction unit is connected with the first position of the two-position four-way valve; the first position and the fourth position of the two-position four-way valve are communicated and connected with the inlet of the separation column array, and any chromatographic column in the separation column array is selected for separation; the outlet of the separation column array is connected with the inlet of the detection unit, the detection unit detects chromatographic signals, and the outlet of the detection unit is connected with the inlet of the mixer through a tee T1 and a tee T2; the diluent pump is connected with the inlet of the mixer through a tee T2, the mixer dilutes the column and then flows out of the sample, and the outlet of the mixer is connected with one end of a two-way valve V2; the two-way valve V2 is conducted and is connected with the third position of the two-position four-way valve through a tee T3; conducting the third position and the second position of the two-position four-way valve, connecting the second position of the two-position four-way valve with a port X of the enrichment column array (the port X of the enrichment column array is the inlet of the enrichment column array at the moment), selecting the enrichment column to enrich the target sample, and connecting a port Y (the port Y of the enrichment column array is the outlet of the enrichment column array at the moment) with a two-way valve V4 through a tee T4; the two-way valve V4 is conducted, and the other end of the two-way valve V4 is output to the waste liquid barrel.

In fig. 9, the two-position four-way valve is in the B state, in which the two-way valves V1 and V3 are in the on state, and the two-way valves V2 and V4 are in the off state. The outlet of the transfusion unit is connected with the inlet of the sample injection unit, and the sample injection unit is in a non-sample injection straight-through state; the outlet of the sample introduction unit is connected with the first position of the two-position four-way valve; the first position of the two-position four-way valve is communicated with the second position of the two-position four-way valve, and the second position of the two-position four-way valve is connected with the interface X of the enrichment column array (at the moment, the interface X of the enrichment column array is the inlet of the enrichment column array); eluting a target sample in the enrichment column, wherein the target sample can be loaded to the separation column or separated through the separation column; an interface Y of the enrichment column array (at the moment, the interface Y of the enrichment column array is an outlet of the enrichment column array) is connected with one end of a two-way valve V3 through a tee T4; the two-way valve V3 is conducted, the other end of the two-way valve V3 is connected with the third position of the two-position four-way valve through the three-way T3, and the third position and the fourth position of the two-position four-way valve are conducted; the fourth position of the two-position four-way valve is connected with the inlet of the liquid chromatographic separation column array; selecting any chromatographic column in the separation column array for separation; the outlet of the separation column array is connected with the inlet of the detection unit, the detection unit detects chromatographic signals, the outlet of the detection unit is connected with one end of a two-way valve V1 through a tee T1, and the other end of the two-way valve V1 is connected with the inlet of a waste liquid barrel or a collector, so that sample collection is realized.

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 three stages of enrichment column arrays 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 27 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 a 27 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 four-way valve in fig. 12(a) is in the a state, and the two-position four-way valve in fig. 12(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 four-way valve is in the state A, see FIG. 2; 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 18 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 19 th to 27 th of the enrichment column array for use in third-dimensional separation; the steps are repeated until enough compounds exist in the 1 st to the 18 th enrichment columns of the enrichment column array, and the control of the second-dimension separation process is carried out.

And (3) controlling a second-dimension loading process: after the control of the first-dimensional separation process is finished, the sample injection unit should be switched to a non-sample injection straight-through state, the two-position four-way valve is switched to a B state, and a second-dimensional chromatographic separation column, for example, a No. 2 separation column, is selected and manually conducted, which is shown in FIG. 3; and selecting one enrichment column from the 1 st enrichment column to the 18 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 completing the sample loading process of the second-dimensional separation.

And (3) controlling a second-dimension separation process: after the second-dimensional sample loading process is completed, when the two-position four-way valve is switched to the state A, the second-dimensional separation is started, and the second-dimensional sample loading process is shown in FIG. 2; in the second dimension separation process, under the assistance of a diluent pump, target fractions are sequentially switched to the 19 th enrichment column to the 27 th enrichment column of the enrichment column array according to sample properties and detection signals for enrichment.

And (3) controlling a third-dimensional separation process: after the control of the second dimension separation process is finished, the two-position four-way valve is switched to the state B, which is shown in figure 3; 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 19 th to 27 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, and a plurality of fractions are sequentially collected at the outlet of the two-way valve V1 by using a collector; and repeating the steps to finish the third-dimensional separation of all samples.

Claims

1. A production type circulating multidimensional liquid chromatography separation system is characterized by comprising a transfusion unit, a sample introduction unit, a detection unit, a separation column array, an enrichment column array, a two-position four-way valve, a diluent pump and a mixer; the detection unit is used for detecting chromatographic signals in the separation process; the sample introduction unit is used for sample introduction; the infusion unit is used for completing the supply of a liquid chromatography separation gradient elution mobile phase;

the separation column array is externally provided with a fixed inlet and a fixed outlet;

the enrichment column array is externally provided with two interfaces, namely an interface X and an interface Y;

the two-position four-way valve is provided with 4 interfaces including a first position, a second position, a third position and a fourth position;

the outlet of the transfusion unit is used for being connected with the inlet of the sample injection unit, and the outlet of the sample injection unit is used for being connected with the first position of the two-position four-way valve;

the fourth position of the two-position four-way valve is used for being connected with the inlet of the separation column array; and the inlet of the detection unit is used for being connected with the outlet of the separation column array.

2. The separation system of claim 1, further comprising two-way valves V1-V4, three-way valves T1-T4, and a connecting line, wherein the connecting line is any one of the following connections (a) to (C):

(A) the outlet of the detection unit is connected with a first interface of a tee T1; a second interface of the tee T1 is connected with one end of a two-way valve V1, and the other end of the two-way valve V1 is output to the waste liquid barrel; a third interface of the tee T1 is connected with a first interface of the tee T2, a second interface of the tee T2 is connected with an outlet of the diluent pump, a third interface of the tee T2 is connected with an inlet of the mixer, and an outlet of the mixer is connected with one end of a two-way valve V2; the other end of the two-way valve V2 is connected with a first interface of a tee T3, a second interface of the tee T3 is connected with one end of a two-way valve V3, and the other end of the two-way valve V3 is connected with the third position of the two-position four-way valve; a third interface of the tee T3 is connected with an interface Y of the enrichment column array, and an interface X of the enrichment column array is connected with a first interface of the tee T4; the second interface of the tee T4 is connected with the second interface of the two-position four-way valve, the third interface of the tee T4 is connected with one end of a two-way valve V4, and the other end of the two-way valve V4 is output to a waste liquid barrel;

(B) the outlet of the detection unit is connected with a first interface of a tee T1; a second interface of the tee T1 is connected with one end of a two-way valve V1, and the other end of the two-way valve V1 is output to the waste liquid barrel; a third interface of the tee T1 is connected with a first interface of the tee T2, a second interface of the tee T2 is connected with an outlet of the diluent pump, a third interface of the tee T2 is connected with an inlet of the mixer, and an outlet of the mixer is connected with one end of a two-way valve V2; the other end of the two-way valve V2 is connected with a first interface of a tee T3, a second interface of the tee T3 is connected with an interface Y of an enrichment column array, and an interface X of the enrichment column array is connected with the second position of the two-position four-way valve; a third interface of the tee T3 is connected with one end of a two-way valve V3, the other end of the two-way valve V3 is connected with a first interface of the tee T4, and a second interface of the tee T4 is connected with a third interface of the two-position four-way valve; a third interface of the tee T4 is connected with one end of a two-way valve V4, and the other end of the two-way valve V4 is output to the waste liquid barrel;

(C) the outlet of the detection unit is connected with a first interface of a tee T1; a second interface of the tee T1 is connected with one end of a two-way valve V1, and the other end of the two-way valve V1 is output to the waste liquid barrel; a third interface of the tee T1 is connected with a first interface of the tee T2, a second interface of the tee T2 is connected with an outlet of the diluent pump, a third interface of the tee T2 is connected with an inlet of the mixer, and an outlet of the mixer is connected with one end of a two-way valve V2; the other end of the two-way valve V2 is connected with a first interface of a tee T3, and a second interface of the tee T3 is connected with a third interface of the two-position four-way valve; a third interface of the tee T3 is connected with one end of a two-way valve V3, and the other end of the two-way valve V3 is connected with a first interface of the tee T4; a second interface of the tee T4 is connected with an interface Y of the enrichment column array, and an interface X of the enrichment column array is connected with the second position of the two-position four-way valve; and a third interface of the tee T4 is connected with one end of a two-way valve V4, and the other end of the two-way valve V4 is output to the waste liquid barrel.