CN113341030A - High-throughput liquid chromatography-mass spectrometry system and separation and analysis method - Google Patents
High-throughput liquid chromatography-mass spectrometry system and separation and analysis method Download PDFInfo
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- 238000004458 analytical method Methods 0.000 title claims abstract description 40
- 238000000926 separation method Methods 0.000 title claims abstract description 30
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 title claims abstract description 18
- 239000000523 sample Substances 0.000 claims abstract description 51
- 238000000605 extraction Methods 0.000 claims description 29
- 239000007788 liquid Substances 0.000 claims description 28
- 239000002699 waste material Substances 0.000 claims description 23
- 238000010828 elution Methods 0.000 claims description 20
- 238000004140 cleaning Methods 0.000 claims description 19
- 239000012071 phase Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 15
- 238000001514 detection method Methods 0.000 claims description 9
- 230000000717 retained effect Effects 0.000 claims description 8
- 238000001802 infusion Methods 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000012535 impurity Substances 0.000 claims description 5
- 238000004949 mass spectrometry Methods 0.000 claims description 5
- 238000001819 mass spectrum Methods 0.000 claims description 5
- 239000003085 diluting agent Substances 0.000 claims description 4
- 239000012488 sample solution Substances 0.000 claims description 4
- 238000012546 transfer Methods 0.000 claims description 4
- 238000002211 ultraviolet spectrum Methods 0.000 claims description 4
- 229910000831 Steel Inorganic materials 0.000 claims description 3
- 239000007791 liquid phase Substances 0.000 claims description 3
- 239000010959 steel Substances 0.000 claims description 3
- 239000012074 organic phase Substances 0.000 claims description 2
- 150000003839 salts Chemical class 0.000 claims description 2
- 230000000712 assembly Effects 0.000 claims 1
- 238000000429 assembly Methods 0.000 claims 1
- 239000012472 biological sample Substances 0.000 abstract description 3
- 239000008280 blood Substances 0.000 abstract description 2
- 210000004369 blood Anatomy 0.000 abstract description 2
- 210000002381 plasma Anatomy 0.000 abstract description 2
- 210000002966 serum Anatomy 0.000 abstract description 2
- 230000002708 enhancing effect Effects 0.000 abstract 1
- 239000000126 substance Substances 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 3
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- 239000007853 buffer solution Substances 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/60—Construction of the column
- G01N30/6034—Construction of the column joining multiple columns
- G01N30/6043—Construction of the column joining multiple columns in parallel
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
Abstract
The invention relates to the field of systems for performing liquid chromatography-mass spectrometry analysis on biological samples such as whole blood, serum, plasma and the like, in particular to a high-throughput liquid chromatography-mass spectrometry system and a separation and analysis method. The invention has two sets of separation and analysis systems which work simultaneously, and can alternately use the sample injector and the capture column, thereby enhancing the utilization rate of the system, greatly saving the analysis time and greatly improving the utilization rate of the mass spectrometer.
Description
Technical Field
The invention relates to the field of systems for performing liquid chromatography-mass spectrometry analysis on biological samples such as whole blood, serum, plasma and the like, in particular to a high-throughput liquid chromatography-mass spectrometry system and a separation and analysis method.
Background
When the mass spectrum is used for measuring the molecular weight of a known target compound, if the impurity content is high or the concentration of a substance to be measured is low, the abundance and the resolution of a corresponding peak of the target compound are low, the interference of impurities is particularly obvious under the condition, and the sample cannot be accurately measured.
In mass spectrometry and detection of complex samples, a sample pretreatment technology is important. In recent years, various sample pretreatment technologies have been developed rapidly, and are suitable for pretreatment of various types of samples. Technologies such as solid phase extraction, solid phase micro-extraction, magnetic extraction, ultrasound assisted extraction, ultrafiltration and dialysis are widely applied to complex biological samples. However, due to the disadvantages of long sample pretreatment time, complicated operation, difficulty in realizing high throughput, automation, etc., researchers have developed various methods to find new techniques to reduce the pretreatment time.
The LC-MS method has obvious advantages in the aspect of processing relatively complex samples, can firstly obtain relatively pure substances to be detected and then carry out mass spectrum detection, can avoid the interference of impurities in the samples to a certain extent, but the traditional LC-MS equipment also has a plurality of problems: 1, nonvolatile inorganic salt can not be used as buffer solution, which can cause inaccurate analysis results of some organic substances which are easy to dissociate in neutral aqueous solution; 2, the mass spectrometer needs to wait for the end of the chromatographic separation and analysis process, so that the utilization rate of the mass spectrometer is low; 3, for the large-flow-rate LC-MS method, shunting treatment is often required before the LC-MS is performed, but the shunting may cause the distribution of the substance to be detected to be uneven, so that an isotope internal standard is required to be added to calibrate the analysis method.
Disclosure of Invention
The invention aims to provide a high-throughput liquid chromatography-mass spectrometry system and a separation and analysis method.
In order to achieve the purpose, the invention adopts the following technical scheme:
the utility model provides a high flux liquid chromatography-mass spectrometry system, includes six transfer pumps, two sets of chromatographic column combination and two detectors to constitute two sets of separation analytic system, two sets separation analytic system finally collects and gets into the mass spectrograph on same root capture column, six the transfer pump is pump one, pump two, pump three, pump four, scavenging pump and elution pump respectively, two sets separation analytic system still includes six sets of six-way valve, column warm box, autosampler module, two extraction columns, two analysis columns and a capture column, six sets of six-way valve is six-way valve A, six-way valve B, six-way valve C, six-way valve D, six-way valve E, six-way valve F respectively, every group six interfaces on the six-way valve are interface 1 department, interface 2 department, interface 3 department, interface 4 department, interface 5 department, interface 6 department respectively.
Furthermore, all parts in the two sets of separation and analysis systems are connected with each other through steel capillaries, the joints of pipelines and the parts are connected through conical joints, and the inverted T-shaped pipelines are connected through tee joints.
Further, the specific connection mode is as follows: an outlet of the first pump is connected to the automatic sample injector module, an outlet of the automatic sample injector module is connected to a tee joint, outlets of the third pump are also connected to the tee joint, the tee joint is connected to a port 4 of a six-way valve E, a port 1 and a port 2 of the six-way valve E are connected with each other, a port 3 and a port 5 of the six-way valve D are connected with each other, a port 5 and a port 2 of the six-way valve E are connected with each other, and a port 6 of the six-way valve E is connected with a cleaning pump; the second pump is connected with a port 5 of the six-way valve F, a port 4 of the six-way valve F is connected with one of the analytical columns, a port 3 of the six-way valve F and a port 6 of the six-way valve F are respectively connected with an inlet and an outlet of one of the extraction columns, and a port 1 of the six-way valve F is connected with waste liquid; the fourth pump is connected with a port 2 of the six-way valve D, a port 3 of the six-way valve D is connected with another analytical column, a port 4 of the six-way valve D and a port 1 of the six-way valve D are respectively connected with an inlet and an outlet of another extraction column, and a port 6 of the six-way valve D is connected with waste liquid; the outlet of one of the analysis columns is connected with a first detector and then connected with a port 2 of the six-way valve C; the outlet of the other analysis column is connected with a second detector, then is connected with a port 4 of the six-way valve C, a port 3 of the six-way valve C is connected with a port 6 of the six-way valve B, the elution pump is connected with a port 2 of the six-way valve B, two ends of the capture column are connected with a port 1 of the six-way valve B and a port 4 of the six-way valve B, a port 3 of the six-way valve B is connected with a port 2 of the six-way valve A, and a port 1 of the six-way valve A is connected with the mass spectrometer; and a joint 3 of the six-way valve A, a joint 5 of the six-way valve B, a joint of the six-way valve C, a joint 6 of the six-way valve D and a joint 1 of the six-way valve F are connected with the waste liquid pool.
Furthermore, two separation systems formed by combining the two sets of chromatographic columns are subjected to sample injection by the same sample injector.
The invention has the beneficial effects that:
the invention comprises liquid phase pump equipment, a valve switching system, an online solid phase extraction system, an ultraviolet detector and mass spectrum detector equipment, and is matched with a modularized automatic sample injector or automatic pretreatment equipment of the same type for use; the device can be controlled by computer software to coordinate, so that two samples can be purified and separated on respective chromatographic columns simultaneously, the same chromatographic column is used for capturing an object to be detected in turn, and finally mass spectrometry is performed; therefore, the analysis time is greatly saved, and the utilization rate of the mass spectrometer is greatly improved.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required to be used in the embodiments of the present invention will be briefly described below.
FIG. 1 is a block flow diagram of the present invention;
FIG. 2 is a block diagram of the connection scheme of the present invention;
fig. 3 is a schematic perspective view of the present invention.
In the figure:
1 pump I, 2 pump II, 3 pump III, 4 pump IV, 5 cleaning pump, 6 elution pump, 7 six-way valve A, 8 six-way valve B, 9 six-way valve C, 10 six-way valve D, 11 six-way valve E, 12 six-way valve F and 13 column temperature box.
Detailed Description
The technical scheme of the invention is further explained by the specific implementation mode in combination with the attached drawings.
Wherein the showings are for the purpose of illustration only and are shown by way of illustration only and not in actual form, and are not to be construed as limiting the present patent; to better illustrate the embodiments of the present invention, some components of the drawings may be omitted, enlarged or reduced, and do not represent the size of an actual product.
Referring to fig. 1, 2 and 3, the high-throughput liquid mass spectrometry system includes six infusion pumps, two sets of chromatography column combinations and two detectors, and two sets of separation analysis systems are formed, the two sets of separation analysis systems are finally collected on the same capture column and enter a mass spectrometer, the six infusion pumps are respectively a pump-one 1, a pump-two 2, a pump-three 3, a pump-four 4, a cleaning pump 5 and an elution pump 6, the two sets of separation analysis systems further include six sets of six-way valves, a column incubator 13, an autosampler module, two extraction columns, two analysis columns and one capture column, the six sets of six-way valves are respectively a six-way valve a7, a six-way valve B8, a six-way valve C9, a six-way valve D10, a six-way valve E11 and a six-way valve F12, and the six interfaces on each set of six-way valves are respectively a port 1, a port 2, a port 3, a port 4, a port 5 and a port 6.
The parts of the two sets of separation and analysis systems are connected by steel capillaries, the joints of the pipelines and the parts are connected by conical joints, and the inverted T-shaped pipelines are connected by a tee joint.
As shown in fig. 2, the specific connection method is as follows: the first pump outlet is connected to the autosampler module, the second autosampler module outlet is connected to a tee joint, the third pump outlet is also connected to the tee joint, the tee joint is connected to the interface 4 of the six-way valve E11, the interface 1 and the interface 2 of the six-way valve E11 are connected with each other, the interface 3 of the six-way valve E11 and the interface 5 of the six-way valve D10 are connected with each other, the interface 5 of the six-way valve E11 and the interface 2 of the six-way valve F12 are connected with each other, and the interface 6 of the six-way valve E11 is connected with the cleaning pump 5; the second pump is connected with a port 5 of the six-way valve F12, a port 4 of the six-way valve F12 is connected with one of the analytical columns, a port 3 of the six-way valve F12 and a port 6 of the six-way valve F12 are respectively connected with an inlet and an outlet of one of the extraction columns, and a port 1 of the six-way valve F12 is connected with waste liquid; the pump four is connected with a port 2 of the six-way valve D10, a port 3 of the six-way valve D10 is connected with another analytical column, a port 4 of the six-way valve D10 and a port 1 of the six-way valve D10 are respectively connected with an inlet and an outlet of another extraction column, and a port 6 of the six-way valve D10 is connected with waste liquid; one of the analytical column outlets is connected with a first detector, and then connected with a port 2 of a six-way valve C9; the outlet of the other analytical column is connected with a second detector, and then is connected with a port 4 of a six-way valve C9, a port 3 of a six-way valve C9 is connected with a port 6 of a six-way valve B8, an elution pump is connected with a port 2 of a six-way valve B8, two ends of the capture column are connected with a port 1 of the six-way valve B8 and a port 4 of a six-way valve B8, a port 3 of the six-way valve B8 is connected with a port 2 of a six-way valve A7, and a port 1 of the six-way valve A7 is connected with a mass spectrometer; the joint 3 of the six-way valve A7, the joint 5 of the six-way valve B8, the joint of the six-way valve C9, the joint 6 of the six-way valve D10 and the joint 1 of the six-way valve F12 are connected with a waste liquid pool.
Wherein, two separation systems formed by combining two sets of chromatographic columns are injected by the same sample injector.
As shown in FIG. 1, a high throughput LC-MS system performs the following steps of separation and analysis of a sample to be tested:
stage one, a sample injection balancing stage:
1. the liquid phase sample injection needle starts to quantitatively absorb a sample solution and temporarily stores the sample solution in the quantitative ring, a mobile phase output by the pump I flows through the quantitative ring from the sample I to the tee joint and is converged with a diluent output by the pump III to the position of an E interface 4 of the six-way valve, and flows to the extraction column I from the position of an E interface 5 of the six-way valve, the position of an F interface 2 of the six-way valve and the position of an F interface 3 of the six-way valve, a component to be detected is retained in the extraction column I, and the mobile phase and impurities which are not retained flow to waste liquid from the position of an F interface 6 of the six-way valve and the position of an F interface 1 of the six-way valve;
2. cleaning liquid output by the cleaning pump washes the second extraction column through a position from an interface E interface 6 of the six-way valve to a position from an interface E interface 1 of the six-way valve to a position from an interface E2 of the six-way valve to a position from an interface E3 of the six-way valve to a position from an interface D5 of the six-way valve to an interface D4 of the six-way valve, and waste liquid is discharged from a position from an interface D1 of the six-way valve to an interface D6 of the six-way valve;
3. the flowing phase output by the pump two passes through a six-way valve F interface 5, a six-way valve F interface 4, an analytical column one, a six-way valve C interface 2, a six-way valve C interface 3, a six-way valve B interface 6 and a six-way valve B interface 1, and flows to waste liquid through a capture column, a six-way valve B interface 4 and a six-way valve B interface 5, which are the balancing processes of the analytical column one and the capture column, and simultaneously the flowing phase output by the pump four passes through a capillary tube, the six-way valve D interface 2, the six-way valve D interface 3, the analytical column two, the detector two, the six-way valve C interface 4, the six-way valve C interface 5 and the six-way valve C interface 1 and flows to waste liquid, which is the balancing process of the analytical column two;
stage two, separation detection stage:
1. the valve starts to be switched, the position of a six-way valve F interface 5 connected with a six-way valve F interface 4 is switched to the position of the six-way valve F interface 5 connected with a six-way valve F interface 6, at the moment, a pump two mobile phase is eluted from the position of the six-way valve F interface 5 to the position of the six-way valve F interface 6 to a first extraction column, a retained sample is cut back to the position of the six-way valve F interface 5 connected with the six-way valve F interface 4 after the elution is finished, the sample enters a detector I after being separated by an analysis column through the position of the six-way valve F interface 3 to the position of the six-way valve F interface 4 to the position of the six-way valve B interface 1 to a capture column, and waste liquid is discharged from the position of the six-way valve B interface 4 to the position of the six-way valve B interface 5 to a waste liquid port;
2. after the valve is switched back to the position of a six-way valve F interface 4 at a six-way valve F interface 5, the position of the six-way valve E interface 5 connected with the six-way valve E interface 4 is switched to the position of the six-way valve E interface 5 connected with the six-way valve E interface 6, cleaning liquid output by the cleaning pump is cleaned and regenerated from the position of the six-way valve E interface 6, the position of the six-way valve E interface 5, the position of the six-way valve F interface 2, the position of the six-way valve F interface 3 and the position of the extraction column I; meanwhile, the mobile phase output by the first pump drives the second sample to be mixed with the diluent output by the third pump at the tee joint, and the second sample passes through a position from an E interface 4 of the six-way valve to a position from an E interface 3 of the six-way valve to a position from a D interface 5 of the six-way valve to a D interface 4 of the six-way valve and then is retained in the well-washed second extraction column;
stage three, sample transfer mass spectrometry detector stage:
1. when a sample two enters the capturing column, the position of a six-way valve B interface 3 connected with a six-way valve B interface 2 is cut into the position of a six-way valve B interface 3 connected with a six-way valve B interface 4, an elution pump firstly outputs a large proportion of water phase from the position of the six-way valve B interface 2 to the position of the six-way valve B interface 1 to the position of the capturing column to the six-way valve B interface 4 to the position of the six-way valve B interface 3 to the position of the six-way valve A interface 2 to the position of the six-way valve A interface 3, salt in the column is washed and discharged by waste liquid, then the six-way valve is switched from the position of the six-way valve A interface 2 connected with the six-way valve A interface 3 to the position of the six-way valve A interface 2, and then the elution pump outputs a pure organic phase to elute the sample two of the capturing column from the position of the six-way valve B interface 2 to the position of the six-way valve B interface 1 to the position of the capturing column to the position of the six-way valve B interface 4 to the position of the six-way valve B interface 3 to the position of the six-way valve A interface 2 to the six-way valve A interface 1, detecting by a mass spectrometer, switching the six-way valve back to a position where a port A2 of the six-way valve is connected with a port A3 of the six-way valve after the detection is finished, outputting a large proportion of water by an elution pump to regenerate the capturing column, switching the six-way valve back to a position where a port B3 of the six-way valve is connected with a port B2 of the six-way valve, switching a port C of the six-way valve back to a position where a port C4 of the six-way valve is connected with a port C3 of the six-way valve, and balancing the capturing column;
2. the position of a six-way valve D interface 3 connected with a six-way valve D interface 2 is switched to the position of a six-way valve D interface 1 connected with the six-way valve D interface 2, the mobile phase output by a pump four is separated from the position of the six-way valve D interface 4 to the position of the six-way valve D interface 3 to the position of an analysis column two through the six-way valve D interface 2 to the position of the six-way valve D interface 1, and then the mobile phase is sent to a detector two to obtain an ultraviolet spectrum, after the ultraviolet spectrum is obtained, the six-way valve is switched back to the position of the six-way valve D interface 2 connected with the six-way valve D interface 3, the position of the six-way valve E interface 3 is connected with the six-way valve E interface 2, the cleaning pump regenerates the extraction column two, meanwhile, a third needle sample starts to be injected, and the first needle flow is repeated; after the second sample comes out of the second detector, the second sample is captured by a balanced capture column from a position of a port C4 of the six-way valve to a position of a port C3 of the six-way valve to a position of a port B6 of the six-way valve to a position of a port B1 of the six-way valve, the six-way valves A and B and the elution pump repeat the process of the step 1, the second sample is sent to a mass spectrum detector for detection, after the detection is finished, the six-way valve is switched back to a position of the port A2 of the six-way valve to be connected with the port A3 of the six-way valve, the elution pump outputs a large proportion of water to regenerate the capture column, and then the six-way valve is switched back to a position of the port B3 of the six-way valve to be connected with the port B2 of the six-way valve;
3. and after the third needle sample is eluted from the second extraction column, the second extraction column is regenerated by the cleaning pump, and meanwhile, the sample injection of the fourth needle sample is started, and the second needle process is repeated until all samples are analyzed.
The foregoing is merely exemplary and illustrative of the present invention and various modifications, additions and substitutions may be made by those skilled in the art to the specific embodiments described without departing from the scope of the invention as defined in the following claims.
Claims (7)
1. A high-flux liquid chromatography-mass spectrometry system is characterized by comprising a plurality of infusion pumps, a chromatographic column assembly and a detector, which form a plurality of separation analysis systems;
the plurality of separation and analysis systems are finally collected on the same capture column and enter a mass spectrometer;
the infusion pumps are respectively a plurality of pumps, a cleaning pump and an elution pump, and the plurality of separation analysis systems further comprise a six-way valve, a column temperature box and an automatic sample injector module;
each separation and analysis system is provided with an extraction column and an analysis column, and finally the whole system is provided with a capture column, and the six-way valve is provided with a corresponding number of interfaces;
and simultaneously and respectively purifying and separating a plurality of samples on respective chromatographic columns, alternately capturing the object to be detected by using the same chromatographic column, and finally performing mass spectrometry.
2. The high throughput LC-MS of claim 1, comprising six infusion pumps, two sets of chromatographic column assemblies and two detectors, and two sets of separation and analysis systems are formed, the two sets of separation and analysis systems are finally converged on the same capture column to enter a mass spectrometer, six infusion pumps are respectively a pump I, a pump II, a pump III, a pump IV, a cleaning pump and an elution pump, the two sets of separation and analysis systems further comprise six groups of six-way valves, a column incubator, an automatic sample injector module, two extraction columns, two analysis columns and one capture column, the six groups of six-way valves are respectively a six-way valve A, a six-way valve B, a six-way valve C, a six-way valve D, a six-way valve E and a six-way valve F, and each group of six interfaces on the six-way valves are respectively an interface 1, an interface 2, an interface 3, an interface 4, an interface 5 and an interface 6.
3. The high-throughput liquid chromatography-mass spectrometry system of claim 1, wherein each of the two separation and analysis systems is connected by a steel capillary, and the connection between the pipeline and the component is connected by a tapered joint, wherein the inverted T-shaped pipeline is connected by a tee.
4. A high throughput liquid chromatography-mass spectrometry system according to claim 1, wherein the specific connection mode is: an outlet of the first pump is connected to the automatic sample injector module, an outlet of the automatic sample injector module is connected to a tee joint, outlets of the third pump are also connected to the tee joint, the tee joint is connected to a port 4 of a six-way valve E, a port 1 and a port 2 of the six-way valve E are connected with each other, a port 3 and a port 5 of the six-way valve D are connected with each other, a port 5 and a port 2 of the six-way valve E are connected with each other, and a port 6 of the six-way valve E is connected with a cleaning pump; the second pump is connected with a port 5 of the six-way valve F, a port 4 of the six-way valve F is connected with one of the analytical columns, a port 3 of the six-way valve F and a port 6 of the six-way valve F are respectively connected with an inlet and an outlet of one of the extraction columns, and a port 1 of the six-way valve F is connected with waste liquid; the fourth pump is connected with a port 2 of the six-way valve D, a port 3 of the six-way valve D is connected with another analytical column, a port 4 of the six-way valve D and a port 1 of the six-way valve D are respectively connected with an inlet and an outlet of another extraction column, and a port 6 of the six-way valve D is connected with waste liquid; the outlet of one of the analysis columns is connected with a first detector and then connected with a port 2 of the six-way valve C; the outlet of the other analysis column is connected with a second detector, then is connected with a port 4 of the six-way valve C, a port 3 of the six-way valve C is connected with a port 6 of the six-way valve B, the elution pump is connected with a port 2 of the six-way valve B, two ends of the capture column are connected with a port 1 of the six-way valve B and a port 4 of the six-way valve B, a port 3 of the six-way valve B is connected with a port 2 of the six-way valve A, and a port 1 of the six-way valve A is connected with the mass spectrometer; and a joint 3 of the six-way valve A, a joint 5 of the six-way valve B, a joint of the six-way valve C, a joint 6 of the six-way valve D and a joint 1 of the six-way valve F are connected with the waste liquid pool.
5. The system according to claim 1, wherein the two separation systems comprising the two sets of chromatographic columns are fed by the same sample feeder.
6. The method for separation and analysis of a high throughput LC-MS system according to any one of claims 2 to 5, wherein:
step one, a sample introduction balance stage:
1. the liquid phase sample injection needle starts to quantitatively absorb a sample solution and temporarily stores the sample solution in the quantitative ring, a mobile phase output by the pump I flows through the quantitative ring from the sample I to the tee joint and is converged with a diluent output by the pump III to the position of an E interface 4 of the six-way valve, and flows to the extraction column I from the position of an E interface 5 of the six-way valve, the position of an F interface 2 of the six-way valve and the position of an F interface 3 of the six-way valve, a component to be detected is retained in the extraction column I, and the mobile phase and impurities which are not retained flow to waste liquid from the position of an F interface 6 of the six-way valve and the position of an F interface 1 of the six-way valve;
2. cleaning liquid output by the cleaning pump washes the second extraction column through a position from an interface E interface 6 of the six-way valve to a position from an interface E interface 1 of the six-way valve to a position from an interface E2 of the six-way valve to a position from an interface E3 of the six-way valve to a position from an interface D5 of the six-way valve to an interface D4 of the six-way valve, and waste liquid is discharged from a position from an interface D1 of the six-way valve to an interface D6 of the six-way valve;
3. the flowing phase output by the pump two passes through a six-way valve F interface 5, a six-way valve F interface 4, an analytical column one, a six-way valve C interface 2, a six-way valve C interface 3, a six-way valve B interface 6 and a six-way valve B interface 1, and flows to waste liquid through a capture column, a six-way valve B interface 4 and a six-way valve B interface 5, which are the balancing processes of the analytical column one and the capture column, and simultaneously the flowing phase output by the pump four passes through a capillary tube, the six-way valve D interface 2, the six-way valve D interface 3, the analytical column two, the detector two, the six-way valve C interface 4, the six-way valve C interface 5 and the six-way valve C interface 1 and flows to waste liquid, which is the balancing process of the analytical column two;
step two, a separation detection stage:
1. the valve starts to be switched, the position of a six-way valve F interface 5 connected with a six-way valve F interface 4 is switched to the position of the six-way valve F interface 5 connected with a six-way valve F interface 6, at the moment, a pump two mobile phase is eluted from the position of the six-way valve F interface 5 to the position of the six-way valve F interface 6 to a first extraction column, a retained sample is cut back to the position of the six-way valve F interface 5 connected with the six-way valve F interface 4 after the elution is finished, the sample enters a detector I after being separated by an analysis column through the position of the six-way valve F interface 3 to the position of the six-way valve F interface 4 to the position of the six-way valve B interface 1 to a capture column, and waste liquid is discharged from the position of the six-way valve B interface 4 to the position of the six-way valve B interface 5 to a waste liquid port;
2. after the valve is switched back to the position of a six-way valve F interface 4 at a six-way valve F interface 5, the position of the six-way valve E interface 5 connected with the six-way valve E interface 4 is switched to the position of the six-way valve E interface 5 connected with the six-way valve E interface 6, cleaning liquid output by the cleaning pump is cleaned and regenerated from the position of the six-way valve E interface 6, the position of the six-way valve E interface 5, the position of the six-way valve F interface 2, the position of the six-way valve F interface 3 and the position of the extraction column I; meanwhile, the mobile phase output by the first pump drives the second sample and the diluent output by the third pump to be mixed at the tee joint, and the second sample passes through a position from an E interface 4 of the six-way valve to a position from an E interface 3 of the six-way valve to a position from a D interface 5 of the six-way valve to a position from the D interface 4 of the six-way valve and then is retained in the well-washed second extraction column.
7. The separation and analysis method according to any one of claim 6, wherein:
further comprising step 3, the sample transfer mass spectrometer stage:
1. when a sample two enters the capturing column, the position of a six-way valve B interface 3 connected with a six-way valve B interface 2 is cut into the position of a six-way valve B interface 3 connected with a six-way valve B interface 4, an elution pump firstly outputs a large proportion of water phase from the position of the six-way valve B interface 2 to the position of the six-way valve B interface 1 to the position of the capturing column to the six-way valve B interface 4 to the position of the six-way valve B interface 3 to the position of the six-way valve A interface 2 to the position of the six-way valve A interface 3, salt in the column is washed and discharged by waste liquid, then the six-way valve is switched from the position of the six-way valve A interface 2 connected with the six-way valve A interface 3 to the position of the six-way valve A interface 2, and then the elution pump outputs a pure organic phase to elute the sample two of the capturing column from the position of the six-way valve B interface 2 to the position of the six-way valve B interface 1 to the position of the capturing column to the position of the six-way valve B interface 4 to the position of the six-way valve B interface 3 to the position of the six-way valve A interface 2 to the six-way valve A interface 1, detecting by a mass spectrometer, switching the six-way valve back to a position where a port A2 of the six-way valve is connected with a port A3 of the six-way valve after the detection is finished, outputting a large proportion of water by an elution pump to regenerate the capturing column, switching the six-way valve back to a position where a port B3 of the six-way valve is connected with a port B2 of the six-way valve, switching a port C of the six-way valve back to a position where a port C4 of the six-way valve is connected with a port C3 of the six-way valve, and balancing the capturing column;
2. the position of a six-way valve D interface 3 connected with a six-way valve D interface 2 is switched to the position of a six-way valve D interface 1 connected with the six-way valve D interface 2, the mobile phase output by a pump four is separated from the position of the six-way valve D interface 4 to the position of the six-way valve D interface 3 to the position of an analysis column two through the six-way valve D interface 2 to the position of the six-way valve D interface 1, and then the mobile phase is sent to a detector two to obtain an ultraviolet spectrum, after the ultraviolet spectrum is obtained, the six-way valve is switched back to the position of the six-way valve D interface 2 connected with the six-way valve D interface 3, the position of the six-way valve E interface 3 is connected with the six-way valve E interface 2, the cleaning pump regenerates the extraction column two, meanwhile, a third needle sample starts to be injected, and the first needle flow is repeated; after the second sample comes out of the second detector, the second sample is captured by a balanced capture column from a position of a port C4 of the six-way valve to a position of a port C3 of the six-way valve to a position of a port B6 of the six-way valve to a position of a port B1 of the six-way valve, the six-way valves A and B and the elution pump repeat the process of the step 1, the second sample is sent to a mass spectrum detector for detection, after the detection is finished, the six-way valve is switched back to a position of the port A2 of the six-way valve to be connected with the port A3 of the six-way valve, the elution pump outputs a large proportion of water to regenerate the capture column, and then the six-way valve is switched back to a position of the port B3 of the six-way valve to be connected with the port B2 of the six-way valve;
3. and after the third needle sample is eluted from the second extraction column, the second extraction column is regenerated by the cleaning pump, and meanwhile, the sample injection of the fourth needle sample is started, and the second needle process is repeated until all samples are analyzed.
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