WO2021103658A1 - Automatic sample injection and analysis device for multiple samples - Google Patents

Abstract

An automatic sample injection and analysis device for multiple samples, relating to the technical field of cell analysis, and being used for solving the technical problem of sample loss. The automatic sample injection and analysis device for multiple samples comprises a sampling device (100) and a liquid chromatography device (200); the sampling device (100) comprises a multi-channel microfluid switching valve (110) connected to the liquid chromatography device (200). By selectively conducting a passage, connected to the liquid chromatography device (200), in the multi-channel microfluid switching valve (110), the conducted passage can be switched after sample injection of a previous sample is completed, so that a sample injection process of a next sample is performed. Therefore, multiple samples can be automatically injected and analyzed online in real time; sample loading obstacles, caused by evaporation, of a continuously flowing sample liquid are avoided, and the technical problem of sample loss is solved.

Description

Multi-sample automatic sampling analysis device

Cross-references to related applications

This application requires the Chinese patent application CN 201911166159.X filed on November 25, 2019 under the title "Multi-sample automatic sampling analysis device" and the title filed on November 25, 2019 under the title "Multi-sample automatic sampling analysis device". The priority of the Chinese patent application CN 201922053302.6 of "Sample Analysis Device", the entire content of the above application is incorporated herein by reference.

Technical field

The present invention relates to the technical field of cell analysis, in particular to a multi-sample automatic sampling analysis device.

Background technique

In recent years, in view of the current hot research directions in the academia such as cell analysis and drug research, microfluidic technology can be used to simulate different organ models in vitro with its remarkable low-cost and high-throughput characteristics; mass spectrometry technology as a powerful analysis The technology is particularly suitable for identifying the structure of biological macromolecules, such as cell secretion factors or drug molecular structures. Therefore, the combination of microfluidic and mass spectrometry technology and the integration of high-sensitivity mass spectrometry analysis technology at the end of a multi-channel microfluidic chip can perform highly parallel online analysis of different biological samples, which is important for a large number of new drug development stages. The drug toxicity screening work has been greatly improved and improved. In addition, the combination of the two technologies has important developments and applications for the separation, processing and detection of complex multi-component samples, such as environmental monitoring, food testing and other fields.

Traditional methods often use a section of fused silica capillary as an interface to connect the outlet of the microfluidic channel with the inlet of the mass spectrometer. This method lacks a high degree of integration and will cause a certain amount of sample loss. Moreover, multi-channel detection relies on manual manual switching, which is time-consuming and labor-intensive, and cannot achieve automated, high-throughput detection. therefore. How to establish a microfluidic-mass spectrometer interface for online, fast and efficient switching of multiple microfluidics has become a big challenge at present.

Summary of the invention

The present invention provides a multi-sample automatic sample injection analysis device, which is used to solve the technical problem of sample loss.

According to the first aspect of the present invention, the present invention provides a multi-sample automatic sampling analysis device, including a sampling device and a liquid chromatography device, the sampling device including the liquid chromatography device connected through an outlet pipeline Multi-channel microfluidic switching valve,

Wherein, the multi-channel microfluidic switching valve has at least two passages, and the passages are selectively conducted with the outlet pipeline.

In one embodiment, the multi-channel microfluidic switching valve includes a first switching valve, and the passage includes a first inlet/outlet provided on the first switching valve;

The first switching valve is also provided with an external port, one end of the external port is selectively connected to one of the first inlet/outlet ports, and the other end of the external port is connected to the outlet pipe through the outlet pipe. The liquid chromatography device is connected.

In an embodiment, the sampling device further includes:

The chip used to carry the sample; and

A second switching valve arranged above the first switching valve, at least two second inlets/outlets are provided on the second switching valve, and the second inlet/outlet is one-to-one with the first inlet/outlet Corresponding settings,

The second inlet/outlet is used to receive the sample input by the chip and input the sample into the first inlet/outlet.

In one embodiment, the first inlet/outlet is provided on the first switching valve at equal intervals along the circumferential direction of the first switching valve, and the second inlet/outlet is along the second switching valve. The circumferentially equidistantly arranged on the second switching valve.

In one embodiment, the first switching valve is rotatably arranged below the second switching valve, and the angle of each rotation of the first switching valve is equal to two adjacent first inlets/outlets. The angle between.

In one embodiment, at least one sample channel is provided in the chip, and the sample channel communicates with the second inlet/outlet corresponding to the sample channel.

In one embodiment, the liquid chromatography device includes:

Mobile phase storage bottle, which is used to store mobile phase;

A detection device, which is used to detect the sample; and

A through valve with at least two ports, a quantitative ring is arranged between the two ports, one of the ports is in communication with the external port or the mobile phase storage bottle, and the other port is connected with the detection device Or the waste liquid bottle is connected.

In an embodiment, the sampling device further includes a micro-syringe pump for transporting samples into the chip.

In one embodiment, the mobile phase storage bottle is connected to the communication valve through an infusion pump.

In one embodiment, the detection device includes a chromatographic column and a detector connected in sequence.

Compared with the prior art, the present invention has the advantage of being selectively conductive through the channel connected to the liquid chromatography device in the multi-channel microfluidic switching valve, and the conductive channel can be switched after the completion of the previous sample injection. In this way, the next sample injection process can be performed, so that multiple samples can be automatically injected and analyzed online in real time. The continuous flow of sample liquid avoids sample loading obstacles caused by evaporation, thereby solving the technical problem of sample loss.

Description of the drawings

Hereinafter, the present invention will be described in more detail based on embodiments and with reference to the drawings.

FIG. 1 is a schematic structural diagram of an automatic sample injection analysis device for multiple samples in an embodiment of the present invention;

2 is a schematic diagram of the structure of a multi-channel microfluidic switching valve in an embodiment of the present invention;

3 is a schematic diagram of the flow path of the first switching valve in the embodiment of the present invention;

Figure 4 is a schematic diagram of the distribution of the ports of the through valve in the embodiment of the present invention;

Figure 5a is a schematic diagram of a sample loading process in an embodiment of the present invention;

Figure 5b is a schematic diagram of a sample injection process in an embodiment of the present invention;

Figures 6a-6f are schematic diagrams of the sample switching process of the present invention.

Reference signs:

100-sampling device; 110-multi-channel microfluidic switching valve;

200-liquid chromatography device; 210-detection device;

1-Micro syringe pump; 2-chip; 3-second switching valve; 4-first switching valve; 5-port valve; 6-waste bottle; 7-mobile phase storage bottle; 8-infusion pump; 9-chromatography Column; 10-detector; 11- connect the pipeline.

Detailed ways

The present invention will be further described below in conjunction with the accompanying drawings.

As shown in FIG. 1, the present invention provides a multi-sample automatic sampling analysis device, including a sampling device 100 and a liquid chromatography device 200. The sampling device 100 includes a multi-sample device connected to the liquid chromatography device 200 through an outlet pipe 11. The channel microfluidic switching valve 110, wherein the multi-channel microfluidic switching valve 110 has at least two passages, and the passages are selectively connected to the outlet pipe 11, so that the sample can be selected to pass through the outlet pipe 11 through a certain passage. It is delivered to the liquid chromatography device 200.

Specifically, the multi-channel microfluidic switching valve 110 includes a first switching valve 4, the passage includes a first inlet/outlet provided on the first switching valve 4, and the first switching valve 4 is also provided with an external port g. One end of g is selectively connected to and conducted with one of the inlet/outlets, and the other end of the external port g is connected to the liquid chromatography device 200 through the outlet pipe 11.

In the following, description will be made by taking the 12 first inlets/outlets provided on the first switching valve 4 as an example.

Among the 12 first inlets/outlets, 6 are the first inlets and the other 6 are the first outlets. As shown in Figure 3, the 6 first inlets are respectively 1', 3', 5', 7', 9'and 11'; the 6 first outlets are respectively 2', 4', 6', 8', 10' and 12'. One end of the external interface g can selectively communicate with any one of the six first inlets.

The above 6 first outlets can be used as temporary sockets for non-detected samples during sampling, that is, the connecting passage between the first inlet 1'and 2'can be used as temporary sockets for non-detected samples during sampling, and the first inlets 3'and 4 The connected path can be used as a temporary connection path for non-test samples during sampling; the connected path between the first inlet 5'and 6'can be used as a temporary connection path for non-detected samples during sampling; the first inlet 7'and 8'are connected The open path can be used as a temporary connection path for non-test samples during sampling; the connected path of the first inlet 9'and 10' can be used as a temporary connection path for non-test samples during sampling, and the first inlet 11' and 12' are connected The path can be used as a temporary connection path for non-test samples during sampling.

Further, the sampling device 100 further includes a chip 2 for carrying a sample and a second switching valve 3 arranged above the first switching valve 4. Wherein, at least two second inlets/outlets are provided on the second switching valve 3, and the second inlets/outlets are arranged in a one-to-one correspondence with the first inlet/outlet, and the second inlet/outlet is used to receive the sample input by the chip 2, and Enter the sample into the first inlet/outlet.

Understandably, the number of second inlets/outlets on the second switching valve 3 may be 6 or more, for example, 12. In addition, the first inlet/outlet is provided on the first switching valve 4 at equal intervals along the circumferential direction of the first switching valve 4, and the second inlet/outlet is provided on the second switching valve 4 at equal intervals along the circumferential direction of the second switching valve 3. Valve 3 is on. As shown in Figures 2 and 3, the first switching valve 4 is divided into 12 equal parts, and each first inlet/outlet is arranged on a dividing line. It is understandable that, in order to have a one-to-one correspondence between the first switching valve 4 and the second switching valve 3, the second switching valve 3 can also be divided into 12 equal parts, wherein each second inlet/outlet is arranged on a bisecting line .

The first switching valve 4 is rotatably arranged below the second switching valve 3, and the angle each time the first switching valve 4 turns is the included angle between two adjacent first inlets/outlets. Since the distances between the first inlets and outlets are equal, after each rotation of the first switching valve 4, each of the first inlets/outlets on it can find the corresponding first inlet/outlet on the second switching valve 3 again. Second import/export.

For example, the first switching valve 4 has 12 first inlets/outlets, and the angle that the first switching valve 4 turns each time is 60°.

At least one sample channel is provided in the chip 2, and the sample channel is connected to the corresponding second inlet/outlet. For example, the chip 2 has 6 sample channels, namely I, II, III, IV, V, and VI six microchannels. Six sample channels are provided on the chip 2, which can cultivate the same cell line in it, thereby ensuring the consistency of the biological environment.

As shown in Figure 2, the 6 second inlets/outlets a, b, c, d, e and f on the second switching valve 3, these 6 second inlets/outlets a, b, c, d, e and f is respectively connected to the six microchannels I, II, III, IV, V and VI mentioned above. The 6 second inlets/outlets are respectively connected with the 6 first inlets/outlets in a one-to-one correspondence, so that the samples in the chip 2 are input into the first inlet/outlet.

As shown in Fig. 6, the entire rotation of the first switching valve 4 is as follows.

As shown in Figure 6a, when the second inlet/outlet a on the second switching valve 3 performs sample loading, the path formed is Ia-1'-g-11, that is, the sample enters the I path and then enters the second inlet/outlet a , And then enter the first inlet 1'and enter the outlet pipeline 11 through the outer interface g.

As shown in Figure 6b, taking the second inlet/outlet a on the second switching valve 3 as a reference when the sample is loaded, and rotating the first switching valve 4 by 60°, the path formed is II-b-3'-g -11, that is, the sample enters the second inlet/outlet b after entering the II passage, and then enters the first inlet 3'and then enters the outlet pipeline 11 through the outer port g.

As shown in Fig. 6c, taking the second inlet/outlet a on the second switching valve 3 as the reference when the sample is loaded, the first switching valve 4 is rotated 120° (that is, rotated twice 60°), the path formed is III-c-5'-g-11, that is, the sample enters the III passage and then enters the second inlet/outlet c, then enters the first inlet 5', and then enters the outlet pipe 11 through the outer port g.

As shown in Figure 6d, taking the second inlet/outlet a on the second switching valve 3 as a reference when the sample is loaded, the first switching valve 4 is rotated 180° (that is, rotated three times 60°), and the formed passage is IV -d-7'-g-11, that is, the sample enters the second inlet/outlet d after entering the IV channel, then enters the first inlet 7'and then enters the outlet pipe 11 through the outer port g.

As shown in Figure 6e, taking the second inlet/outlet a on the second switching valve 3 as a reference when the sample is loaded, the first switching valve 4 is rotated 240° (that is, rotated four times 60°), and the path formed is Ve-9'-g-11, that is, the sample enters the second inlet/outlet e after entering the V path, then enters the first inlet 9'and then enters the outlet pipe 11 through the outer port g.

As shown in Figure 6f, taking the second inlet/outlet a on the second switching valve 3 as the reference when the sample is loaded, and the first switching valve 4 is rotated 300° (that is, rotated five times 60°), the path formed is VI-f-11'-g-11, that is, the sample enters the VI channel and then enters the second inlet/outlet f, then enters the first inlet 11' and then enters the outlet pipeline 11 through the external interface g.

In addition, the sampling device 100 also includes a micro-syringe pump 1 for delivering samples into the chip 2. Among them, the micro syringe pump 1 can be a Haval syringe pump. The inlets of the sample channels of the chip 2 are respectively connected to the micro-injection pump 1, and the outlets are respectively connected to the six second inlets. The Harvard Syringe Pump continuously delivers gradient concentrations of drug solutions to the 6 microchannels of the chip 2 at a constant speed to form a uniform pumping condition, thereby ensuring a high degree of parallelism, and the continuous flow of the sample solution can avoid evaporation. The sample loading barrier.

The working process of the sampling device 100 will be described in detail below.

As shown in Figure 2, the second inlet/outlet a, b, c, d, e, and f on the second switching valve 3 are respectively connected to the outlets of the six microchannels I, II, III, IV, V, and VI. ; The six first inlets 1', 3', 5', 7', 9'and 11' on the first switching valve 4 are connected to a, b, c, on the second switching valve 3, respectively The six ports d, e and f correspond one-to-one and are in a connected state. The sample liquid enters the first switching valve 4 through the second inlet/outlet a, b, c, d, e, and f on the second switching valve 3. The first inlets 1', 3', 5', 7', 9', and 11' are used to temporarily accept non-detection sample liquid during sampling.

The external port g can selectively communicate with the first inlets 1', 3', 5', 7', 9', and 11' of the first switching valve 4 through the internal flow path. For example, the external port g communicates with the first inlet 1'of the first switching valve 4 through the internal flow path 1'-g, and the sample liquid in the first inlet 1'is output to the liquid chromatography device 200 through the external port g.

Take the analysis of the sample in the I channel connected to the second inlet/outlet a on the second switching valve 3 as an example for description. As shown in Figure 5, the micro-injection pump 1 provides the power required for the entire flow path, and the sample solution flows from the inlet of the chip 2 into the channel between the chip 2 and the second switching valve 3 by the pumping of the micro-injection pump 1 and flows in parallel. Through the entire channel, after flowing out of the channel outlet, it enters the second inlet/outlet a of the second switching valve 3, and then enters the first inlet 1'of the first switching valve 4, passing through the internal flow path 1'-g, and passing through the external interface g And the outgoing pipeline 11 is delivered to the liquid chromatography device 200.

The liquid chromatography device 200 will be described in detail below. The liquid chromatography device 200 includes a mobile phase storage bottle 7, a detection device 210, and a through valve 5 with at least two interfaces. Among them, the mobile phase storage bottle 7 is used to store the mobile phase; the detection device 210 is used to detect samples; a quantitative ring is provided between the two ports of the through valve 5, one of which is connected to the external port g or the mobile phase storage bottle 7 The other interface is connected with the detection device 210 or the waste liquid bottle 6.

As shown in Fig. 4, the description is made by taking the through valve 5 with 6 ports as an example. The 6 ports of the port valve 5 are A, B, C, D, E, and F respectively. Among them, there is a quantitative loop between the interface C and the interface F, which can realize quantitative detection.

The on-off between the six ports of the valve 5 can be selectively switched. For example, when loading samples, interfaces B, C, F and A are connected, and interfaces D and E are connected. In order to make the sample enter the quantitative loop through the external port g and the outgoing pipe 11; when the sample is injected, the ports D, C, F and E are connected, and the ports A and B are connected, so that the mobile phase can pass through the quantitative loop , And the sample in the quantitative loop is brought into the detection device 210.

Specifically, when the sample is loaded, the ports B, C, F, and A are connected, the port B is connected to the outlet pipe 11, and the port A is connected to the waste bottle 6. The sample is transported to the port B through the external port g and the outlet pipe 11, and flows through the quantitative ring between the port C and the port F, and then flows into the waste liquid bottle 6.

When the sample is loaded, the sample is injected. Switch the connected ports in the through valve 5 to connect ports D, C, F and E. The mobile phase in the mobile phase storage bottle 7 enters port D and is transported to the quantitative ring between port C and port F, and drives The sample in the quantitative loop enters the detection device 210 through the interface E for detection.

After the sample injection is completed, the connected ports in the through valve 5 are switched again to connect the ports B, C, F, and A, that is, the path state of the sample loading process is restored.

In addition, the mobile phase storage bottle 7 is connected to the through valve 5 through the infusion pump 8. The infusion pump 8 provides power to the flow path.

The detection device 210 includes a chromatographic column 9 and a detector 10 connected in sequence. The detector can be replaced, so it can be applied to detection items such as the separation of multi-component samples.

The working process of the multi-sample automatic sampling analysis device of the present invention will be described in detail below.

The first step is to load the sample.

As shown in Figure 5a, the micro-injection pump 1 provides the power required for the entire flow path, and the sample solution flows from the inlet of the chip 2 into the channel between the chip 2 and the second switching valve 3 by the pumping of the micro-injection pump 1 and flows in parallel. Through the entire channel, after flowing out of the channel outlet, it enters the second inlet/outlet a of the second switching valve 3, and then enters the first inlet 1'of the first switching valve 4, passing through the internal flow path 1'-g, and passing through the external interface g And the outgoing pipeline 11 is delivered to the liquid chromatography device 200.

The ports B, C, F, and A of the through valve 5 are connected, and the port B is connected to the outlet pipe 11, and the port A is connected to the waste liquid bottle 6. The sample is transported to the port B through the external port g and the outlet pipe 11, and flows through the quantitative ring between the port C and the port F, and then flows into the waste liquid bottle 6.

In the second step, the sample is injected.

As shown in Figure 5b, switch the connected ports in the through valve 5 to connect ports D, C, F and E, then the mobile phase in the mobile phase storage bottle 7 enters port D and is transported between port C and port F And drive the sample in the quantitative ring to enter the chromatographic column 9 and the detector 10 through the interface E for detection.

In the third step, the connected ports in the through valve 5 are switched again, so that the ports B, C, F and A are connected, that is, the path state of the sample loading process is restored.

In the fourth step, the multi-microfluidic switching valve 110 automatically switches to the next sample for detection.

Specifically, the first switching valve 4 is rotated clockwise by 60° as a whole. At this time, the 1'-g channel is switched to the 3'-g channel, and the sample liquid in channel II flows through the second inlet/outlet through b and After the 3'-g passage, it enters the outlet pipe 11, and is then pumped to the port B of the through valve 5.

Repeat the second step of loading and the third step of the sample injection process until the sample in channel II on the chip 2 is detected.

Subsequently, the entire first switching valve 4 is rotated clockwise by 120°, 180°, 240°, and 300° (using the state of the first switching valve 4 when the sample in channel I is detected as the basic position of rotation), correspondingly 1 The'-g channel can be flexibly switched to 5'-g, 7'-g, 9'-g, 11'-g channels to detect samples in channels Ⅲ, Ⅳ, Ⅴ, and Ⅵ on the chip 2 respectively . Thereby, the waste of human resources is greatly reduced, and the detection efficiency is greatly improved.

In summary, the present invention can replace the manual sample change module when the multi-channel chip is coupled with the LC/MS instrument in the prior art, but real-time sampling by mechanically controlling the microfluidic switching valve 110 and the through valve 5. Sampling can reduce the use of human resources, reduce the cost of testing, and obtain a large amount of real-time data when unattended, thereby greatly improving efficiency and realizing high-throughput testing.

It should be noted that in the drawings of the present invention, two components connected by solid lines are in communication with each other.

Although the present invention has been described with reference to the preferred embodiments, without departing from the scope of the present invention, various modifications can be made thereto and the components therein can be replaced with equivalents. In particular, as long as there is no structural conflict, the various technical features mentioned in the various embodiments can be combined in any manner. The present invention is not limited to the specific embodiments disclosed in the text, but includes all technical solutions falling within the scope of the claims.

Claims (10)

1. A multi-sample automatic sampling analysis device, which is characterized in that it comprises a sampling device and a liquid chromatography device. The sampling device comprises a multi-channel microfluidic switching valve connected to the liquid chromatography device through an outlet pipe,

   Wherein, the multi-channel microfluidic switching valve has at least two passages, and the passages are selectively conducted with the outlet pipeline.
2. The multi-sample automatic sample injection analysis device according to claim 1, wherein the multi-channel microfluidic switching valve comprises a first switching valve, and the passage comprises a first switching valve arranged on the first switching valve. import and export;

   The first switching valve is also provided with an external port, one end of the external port is selectively connected to one of the first inlet/outlet ports, and the other end of the external port is connected to the outlet pipe through the outlet pipe. The liquid chromatography device is connected.
3. The multi-sample automatic sampling analysis device according to claim 2, wherein the sampling device further comprises:

   The chip used to carry the sample; and

   A second switching valve arranged above the first switching valve, at least two second inlets/outlets are provided on the second switching valve, and the second inlet/outlet is one-to-one with the first inlet/outlet Corresponding settings,

   The second inlet/outlet is used to receive the sample input by the chip and input the sample into the first inlet/outlet.
4. The multi-sample automatic sampling analysis device according to claim 3, wherein the first inlet/outlet is arranged on the first switching valve at equal intervals along the circumferential direction of the first switching valve, The second inlet/outlet is provided on the second switching valve at equal intervals along the circumferential direction of the second switching valve.
5. The multi-sample automatic sampling analysis device according to claim 4, wherein the first switching valve is rotatably arranged below the second switching valve, and each time the first switching valve is turned The angle of is the angle between two adjacent first inlets/outlets.
6. The multi-sample automatic sample injection analysis device according to any one of claims 3-5, wherein at least one sample channel is provided in the chip, and the sample channel and the second input channel corresponding to it are provided with at least one sample channel. /Exit is connected.
7. The multi-sample automatic sampling analysis device according to any one of claims 2-5, wherein the liquid chromatography device comprises:

   Mobile phase storage bottle, which is used to store mobile phase;

   A detection device, which is used to detect the sample; and

   A through valve with at least two ports, a quantitative ring is arranged between the two ports, one of the ports is in communication with the external port or the mobile phase storage bottle, and the other port is connected with the detection device Or the waste liquid bottle is connected.
8. The multi-sample automatic sample injection analysis device according to any one of claims 3 to 5, wherein the sampling device further comprises a micro-syringe pump for transporting samples into the chip.
9. The multi-sample automatic sampling analysis device according to claim 7, wherein the mobile phase storage bottle is connected to the through valve through an infusion pump.
10. The multi-sample automatic sampling analysis device according to claim 7, wherein the detection device comprises a chromatographic column and a detector connected in sequence.

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