Two-dimensional liquid chromatography microfluidic chip and manufacturing method thereof

Abstract

The application relates to the technical field of substance separation and detection, and relates to a two-dimensional liquid chromatography micro-fluidic chip and a manufacturing method thereof. The two-dimensional liquid chromatography micro-fluidic chip comprises a chip body, and a channel structure is formed in the chip body; a sample channel is connected with a first end of a one-dimensional cross injection channel, a one-dimensional mobile phase channel is connected with a second end of the one-dimensional cross injection channel, a first end of a one-dimensional separation channel is connected with a third end of the one-dimensional cross injection channel, a plunger channel is connected with the third end of the one-dimensional cross injection channel, a two-dimensional mobile phase channel is connected with a first end of a two-dimensional cross injection channel, a second end of the one-dimensional separation channel is connected with a second end of the two-dimensional cross injection channel, a two-dimensional separation channel is connected with a third end of the two-dimensional cross injection channel, and a post-column channel is connected with the two-dimensional separation channel. The two-dimensional liquid chromatography micro-fluidic chip and the manufacturing method thereof provided by the application solve the defect that it is difficult to fill a chromatographic column in a channel of a quartz chip in the prior art.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of substance separation and detection, and particularly relates to a two-dimensional liquid chromatography microfluidic chip and a manufacturing method thereof. BACKGROUND

[0002] For the separation and detection of some complex mixtures, very strong separation capacity is usually required, and traditional one-dimensional separation methods are difficult to meet the separation requirements of such compounds (such as macromolecular proteins and polypeptides). At present, in order to realize greater peak capacity and higher separation efficiency, multi-dimensional separation modes are widely used. Among them, two-dimensional liquid chromatography has become an effective method for separating complex samples due to its excellent separation capacity and peak capacity. However, the traditional two-dimensional liquid chromatography system usually relies on valve switching to realize, which not only makes the system structure complex, but also introduces a large dead volume, so as to fail to meet the separation requirements of precious small-volume samples.

[0003] In comparison, microfluidic chip chromatographic analysis technology exhibits significant advantages. It can greatly reduce the consumption of precious samples, significantly shorten the analysis time, and has the characteristics of flexible design, adaptation to small-volume rare samples, high-efficiency and rapid separation, and convenience for miniaturization and portability. Quartz microfluidic chips have become a commonly used preparation material for liquid chromatography chips due to their excellent pressure resistance and good repeatability. However, due to the greater rigidity of the quartz chip, it is difficult to fill the chromatographic column in the channel of the quartz chip in the prior art. SUMMARY

[0004] The present application provides a two-dimensional liquid chromatography microfluidic chip and a manufacturing method thereof, to solve the defect that it is difficult to fill the chromatographic column in the channel of the quartz chip in the prior art.

[0005] In one aspect, the present application provides a two-dimensional liquid chromatography microfluidic chip, comprising: a chip body, a channel structure is formed in the chip body, and the channel structure comprises a sample channel, a one-dimensional mobile phase channel, a two-dimensional mobile phase channel, a one-dimensional cross-sample injection channel, a two-dimensional cross-sample injection channel, a one-dimensional separation channel, a two-dimensional separation channel, a plunger channel and a post-column channel.

[0006] The sample channel is connected with the first end of the one-dimensional cross-sample injection channel, the one-dimensional mobile phase channel is connected with the second end of the one-dimensional cross-sample injection channel, the first end of the one-dimensional separation channel is connected with the third end of the one-dimensional cross-sample injection channel, the plunger channel is connected with the third end of the one-dimensional cross-sample injection channel, the two-dimensional mobile phase channel is connected with the first end of the two-dimensional cross-sample injection channel, the second end of the one-dimensional separation channel is connected with the second end of the two-dimensional cross-sample injection channel, the first end of the two-dimensional separation channel is connected with the third end of the two-dimensional cross-sample injection channel, and one end of the post-column channel is connected with the second end of the two-dimensional separation channel.

[0007] The two-dimensional liquid chromatography microfluidic chip provided by the application is characterized in that: a first plunger is arranged at the connection position of the plunger channel and the first end of the one-dimensional separation channel; and a second plunger is arranged at the connection position of the two-dimensional mobile phase channel and the first end of the two-dimensional separation channel.

[0008] The two-dimensional liquid chromatography microfluidic chip provided by the application is characterized in that: a first limiting channel for limiting the first plunger is arranged at the third end outlet of the one-dimensional cross-injection channel; and a second limiting channel for limiting the second plunger is arranged at the fourth end inlet of the two-dimensional cross-injection channel.

[0009] The two-dimensional liquid chromatography microfluidic chip provided by the application is characterized in that: the cross sections of the first limiting channel and the second limiting channel are both semicircular.

[0010] The two-dimensional liquid chromatography microfluidic chip provided by the application is characterized in that: the diameters of the first limiting channel and the second limiting channel are both 80 μm to 120 μm.

[0011] The two-dimensional liquid chromatography microfluidic chip provided by the application is characterized in that: the first plunger and the second plunger are both porous silica microspheres.

[0012] The two-dimensional liquid chromatography microfluidic chip provided by the application is characterized in that: the diameters of the one-dimensional separation channel and the two-dimensional separation channel are both 80 μm to 120 μm.

[0013] The two-dimensional liquid chromatography microfluidic chip provided by the application is characterized in that: the channel structure further comprises a first liquid discharge channel and a second liquid discharge channel; the first liquid discharge channel is connected with the fourth end of the one-dimensional cross-injection channel; and the second liquid discharge channel is connected with the fourth end of the two-dimensional cross-injection channel.

[0014] The two-dimensional liquid chromatography microfluidic chip provided by the application is characterized in that: the chip body comprises two quartz monoliths, the quartz monoliths are provided with grooves for forming the channel structure, and the two quartz monoliths are oppositely arranged and bonded to each other, so that the grooves on the two quartz monoliths form the channel structure.

[0015] Another aspect of the application provides a two-dimensional liquid chromatography microfluidic chip and a manufacturing method thereof, comprising the following steps.

[0016] Grooves corresponding to the channel structure are respectively manufactured on the two quartz monoliths.

[0017] The two quartz monoliths with the channel are aligned and heat bonded to the chip body, and the channels of the two quartz monoliths are combined to form the channel structure.

[0018] A first plunger is introduced into the plunger channel through the plunger channel and to the connection between the plunger channel and the first end of the one-dimensional separation channel, and then the one-dimensional separation channel is filled with chromatographic packing through the post-column channel to form a one-dimensional chromatographic packing column in the one-dimensional separation channel.

[0019] The two-dimensional liquid chromatography microfluidic chip provided by the present application can quickly and simply realize the preparation of the one-dimensional chromatographic packing column and the two-dimensional chromatographic packing column in the chip body, and solves the defect that it is difficult to fill the chromatographic column in the channel of the quartz chip in the prior art.

[0020] Additional aspects and advantages of the application will be set forth in part in the description which follows, and in part will become apparent to those skilled in the art upon examination of the following description and drawings. BRIEF DESCRIPTION OF DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the present application or prior art, the following will briefly introduce the drawings needed to be used in the embodiments or prior art description. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without any creative effort on the basis of these drawings.

[0022] Figure 1 is one of the schematic diagrams of the two-dimensional liquid chromatography microfluidic chip provided by the embodiments of the present application.

[0023] Figure 2 is a heat bonding schematic diagram of two quartz monoliths with channel structures in the two-dimensional liquid chromatography microfluidic chip provided by the embodiments of the present application.

[0024] Figure 3 is a schematic diagram of a circular cross-section channel in a two-dimensional liquid chromatography microfluidic chip provided by an embodiment of the present application.

[0025] Figure 4 is a schematic diagram of a semi-circular cross-section channel in a two-dimensional liquid chromatography microfluidic chip provided by an embodiment of the present application.

[0026] Figure 5 is a flowchart of a microfluidic chip manufacturing method provided by an embodiment of the present application.

[0027] Reference signs:

[0028] 100, chip body; 101, sample channel; 102, one-dimensional mobile phase channel; 103, two-dimensional mobile phase channel; 104, one-dimensional cross-injection channel; 105, two-dimensional cross-injection channel; 106, one-dimensional separation channel; 107, two-dimensional separation channel; 108, plunger channel; 109, post-column channel; 110, first limiting channel; 111, second limiting channel; 112, first liquid discharge channel; 113, second liquid discharge channel. DETAILED DESCRIPTION

[0029] In order to make the objectives, technical solutions and advantages of the present application clearer, the technical solutions in the present application will be described clearly and completely below with reference to the drawings in the present application. Obviously, the described embodiments are some of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the scope of protection of the present application.

[0030] In the description of the embodiments of the present application, it should be noted that the terms “center”, “longitudinal”, “transverse”, “upper”, “lower”, “front”, “rear”, “left”, “right”, “vertical”, “horizontal”, “top”, “bottom”, “inner”, “outer” and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the embodiments of the present application and simplifying the description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the embodiments of the present application. In addition, the terms “first”, “second”, “third” are only for the purpose of description, and cannot be understood as indicating or implying relative importance.

[0031] In the description of the embodiments of the present application, it should be noted that unless specifically defined and limited, the terms "connected", "connected to", "connection" should be interpreted broadly, for example, can be fixed connection, can also be detachable connection, or integral connection, can be mechanical connection, can also be electrical connection, can be direct connection, or indirect connection through intermediate medium. For those skilled in the art, the specific meaning of the above terms in the embodiments of the present application can be understood according to the specific circumstances.

[0032] In the embodiments of the present application, unless specifically defined and limited, the first feature is "on" or "under" the second feature, which can be direct contact between the first and second features, or indirect contact between the first and second features through an intermediate medium. Moreover, the first feature "above", "over" and "on" the second feature can be that the first feature is directly above or obliquely above the second feature, or only indicates that the horizontal height of the first feature is higher than that of the second feature. The first feature "below", "under" and "under" the second feature can be that the first feature is directly below or obliquely below the second feature, or only indicates that the horizontal height of the first feature is less than that of the second feature.

[0033] In the description of the present application, the description of the terms "one embodiment", "some embodiments", "example", "specific example" or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present application. In the present application, the illustrative description of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any suitable manner in any one or more embodiments or examples. In addition, those skilled in the art can combine and combine different embodiments or examples described in the present application and the features of different embodiments or examples without contradiction.

[0034] The following will be described in conjunction with Figures 1 to 5 The two-dimensional liquid chromatography microfluidic chip and the manufacturing method thereof are described.

[0035] Referring to Figure 1 As shown in the figure, the two-dimensional liquid chromatography microfluidic chip provided by the embodiments of the present application comprises: a chip body 100, a channel structure is formed in the chip body 100, and the channel structure comprises a sample channel 101, a one-dimensional mobile phase channel 102, a two-dimensional mobile phase channel 103, a one-dimensional cross sample injection channel 104, a two-dimensional cross sample injection channel 105, a one-dimensional separation channel 106, a two-dimensional separation channel 107, a plunger channel 108 and a post-column channel 109.

[0036] The sample channel 101 is connected with the first end of the one-dimensional cross-injection channel 104, the one-dimensional mobile phase channel 102 is connected with the second end of the one-dimensional cross-injection channel 104, the first end of the one-dimensional separation channel 106 is connected with the third end of the one-dimensional cross-injection channel 104, the plunger channel 108 is connected with the third end of the one-dimensional cross-injection channel 104, the two-dimensional mobile phase channel 103 is connected with the first end of the two-dimensional cross-injection channel 105, the second end of the one-dimensional separation channel 106 is connected with the second end of the two-dimensional cross-injection channel 105, the first end of the two-dimensional separation channel 107 is connected with the third end of the two-dimensional cross-injection channel 105, and one end of the post-column channel 109 is connected with the second end of the two-dimensional separation channel 107.

[0037] It should be noted that the first end to the fourth end of the one-dimensional cross-injection channel 104 in the embodiment are the ① to ④ ends shown in the Figure 1 Similarly, the first end to the fourth end of the two-dimensional cross-injection channel 105 in the embodiment are the ① to ④ ends shown in the Figure 1

[0038] The two-dimensional liquid chromatography microfluidic chip provided by the application can quickly and simply realize the preparation of the one-dimensional chromatographic packed column and the two-dimensional chromatographic packed column in the chip body 100, and solves the defect that it is difficult to fill the chromatographic column in the channel of the quartz chip in the prior art.

[0039] Specifically, in the embodiment, the channel structure includes the sample channel 101, the one-dimensional mobile phase channel 102, the two-dimensional mobile phase channel 103, the one-dimensional cross-injection channel 104, the two-dimensional cross-injection channel 105, the one-dimensional separation channel 106, the two-dimensional separation channel 107, the plunger channel 108, and the post-column channel 109.

[0040] ​The sample channel 101 is used for inputting sample solution; the one-dimensional flow phase channel 102 is used for one-dimensional flow phase inflow; the two-dimensional flow phase channel 103 is used for two-dimensional flow phase inflow and is used for introducing a second plunger to the connection between the two-dimensional flow phase channel 103 and the first end of the two-dimensional separation channel 107, so as to prepare a one-dimensional chromatographic packing column and a two-dimensional chromatographic packing column in the one-dimensional separation channel 106 and the two-dimensional separation channel 107 respectively.

[0041] In the one-dimensional cross sample injection channel 104, after the solvent and the sample form a stable laminar flow, the sample is injected in a small volume by entering the one-dimensional separation channel 106; in the two-dimensional cross sample injection channel 105, the fraction flowing out of the one-dimensional separation channel 106 and the two-dimensional flow phase solvent enter the two-dimensional separation channel 107.

[0042] The one-dimensional separation channel 106 is used for packing a one-dimensional chromatographic packing material and is used for one-dimensional separation of the sample; the two-dimensional separation channel 107 is used for packing a two-dimensional chromatographic packing material and is used for two-dimensional separation of the sample.

[0043] The plunger channel 108 is used for introducing a first plunger to the connection between the plunger channel 108 and the first end of the one-dimensional separation channel 106.

[0044] The post-column channel 109 is used for packing chromatographic packing materials in the one-dimensional separation channel 106 and the two-dimensional separation channel 107 and is used for sample solution outflow.

[0045] In use, in the channel structure, the sample channel 101, the one-dimensional flow phase channel 102, the two-dimensional flow phase channel 103, the post-column channel 109, and the following first liquid discharge channel 112 and second liquid discharge channel 113 are all connected with capillary tubes (not shown in the figure), the capillary tubes are used for communicating with corresponding containers outside, so as to facilitate introduction of corresponding liquid media into the channel structure, and valves can be arranged on the capillary tubes, so as to open or close the corresponding channels.

[0046] According to some embodiments of the present application, the connection between the plunger channel 108 and the first end of the one-dimensional separation channel 106 is provided with a first plunger (not shown in the figure), the connection between the two-dimensional flow phase channel 103 and the first end of the two-dimensional separation channel 107 is provided with a second plunger (not shown in the figure); the one-dimensional separation channel 106 is provided with a one-dimensional chromatographic packing column (not shown in the figure), and the two-dimensional separation channel 107 is provided with a two-dimensional chromatographic packing column (not shown in the figure).

[0047] By setting the first plunger and the second plunger in the channel structure, the plunger can be used to block the response position of the channel structure, which facilitates filling of the one-dimensional chromatographic packing column and the two-dimensional chromatographic packing column in the one-dimensional separation channel 106 and the two-dimensional separation channel 107 respectively, and after the one-dimensional chromatographic packing column and the two-dimensional chromatographic packing column are formed in the channel structure, the one-dimensional chromatographic packing column and the two-dimensional chromatographic packing column can be positioned to ensure the stability of the one-dimensional chromatographic packing column and the two-dimensional chromatographic packing column.

[0048] It should be noted that the first plunger is the inlet end plunger of the one-dimensional chromatographic packing column, and the second plunger is the end plunger of the one-dimensional chromatographic packing column and the inlet end plunger of the two-dimensional chromatographic packing column. In addition, the first plunger and the second plunger in the embodiment are both porous plunger structures, which can allow liquid medium to flow.

[0049] Referring to Figure 1 As shown in the drawings, according to some embodiments of the present application, the third end outlet of the one-dimensional cross-injection channel 104 is provided with a first limiting channel 110 for limiting the first plunger, and the fourth end inlet of the two-dimensional cross-injection channel 105 is provided with a second limiting channel 111 for limiting the second plunger.

[0050] By setting the first limiting channel 110 and the second limiting channel 111, the first plunger and the second plunger in the channel structure can be limited respectively under the premise of ensuring the smooth flow of the liquid medium, so as to prevent the first plunger and the second plunger from moving in the channel structure.

[0051] Referring to Figure 3 and Figure 4 As shown in the drawings, according to some embodiments of the present application, the cross sections of the first limiting channel 110 and the second limiting channel 111 are both semicircular.

[0052] By setting the diameters of the first limiting channel 110 and the second limiting channel 111 to be 80 μm to 120 μm, and setting the diameters of the one-dimensional separation channel 106 and the two-dimensional separation channel 107 to be 80 μm to 120 μm, the first plunger and the second plunger can be limited by the solid structure part at the lower part of the semicircular channel, and the liquid medium can also flow smoothly through the semicircular channel.

[0053] Specifically, the cross sections of the first limiting channel 110 and the second limiting channel 111 are both semicircular, the cross sections of the one-dimensional separation channel 106 and the two-dimensional separation channel 107 are both circular, the diameters of the first limiting channel 110 and the second limiting channel 111 can be 80 μm, 90 μm, 100 μm, 110 μm or 120 μm, etc., and the diameters of the one-dimensional separation channel 106 and the two-dimensional separation channel 107 can be 80 μm, 90 μm, 100 μm, 110 μm or 120 μm, etc.

[0054] According to some embodiments of the present invention, both the first plunger and the second plunger are porous silica microspheres.

[0055] By using porous silica microspheres for both the first and second plungers, the microspheres can be used to confine the chromatographic column and provide flow channels for the liquid medium through their porous structure. Furthermore, the porous silica microspheres have a simple structure, are easily introduced into the corresponding positions of the channel structure through the plunger channels, and are stable, not easily reacting with the liquid medium or chromatographic packing material, exhibiting good stability and durability.

[0056] See Figure 1 As shown, according to some embodiments of the present invention, the channel structure further includes a first drainage channel 112 and a second drainage channel 113. The first drainage channel 112 is connected to the fourth end of the one-dimensional cross-shaped injection channel 104, and the second drainage channel 113 is connected to the fourth end of the two-dimensional cross-shaped injection channel 105.

[0057] By setting up the first drainage channel 112 and the second drainage channel 113, the liquid medium at the corresponding position in the channel structure can be discharged.

[0058] See Figure 2 As shown, according to some embodiments of the present invention, the chip body 100 includes two quartz wafers, each quartz wafer having channels for forming a channel structure. The two quartz wafers are arranged opposite to each other and bonded together, so that the channels on the two quartz wafers form a channel structure.

[0059] By setting the chip body 100 as two opposing and bonded quartz wafers, it is easy to create a channel structure between the two quartz wafers.

[0060] The following describes the microfluidic chip fabrication method provided by the present invention. The microfluidic chip fabrication method described below can be referred to in correspondence with the two-dimensional liquid chromatography microfluidic chip described above.

[0061] See Figure 5 As shown, the microfluidic chip fabrication method provided in this embodiment of the invention is used to fabricate a two-dimensional liquid chromatography microfluidic chip as described in any of the above embodiments, and includes the following steps.

[0062] S510. Create channels corresponding to the channel structure on two quartz wafers respectively.

[0063] S520: Align two quartz wafers with channels and thermally bond them to form a chip body 100, so that the channels on the two quartz wafers combine to form a channel structure.

[0064] S530, introducing a first plunger into the plunger channel 108 to the junction of the plunger channel 108 and the first end of the one-dimensional separation channel 106, and then filling the one-dimensional separation channel 106 with chromatographic packing material through the post-column channel 109 to form a one-dimensional chromatographic packing column in the one-dimensional separation channel 106, introducing a second plunger into the two-dimensional mobile phase channel 103 to the junction of the two-dimensional mobile phase channel 103 and the first end of the two-dimensional separation channel 107, and then filling the two-dimensional separation channel 107 with chromatographic packing material through the post-column channel 109 to form a two-dimensional chromatographic packing column in the two-dimensional separation channel 107.

[0065] The microfluidic chip manufacturing method provided by the application can facilitate filling of one-dimensional chromatographic packing columns and two-dimensional chromatographic packing columns in the one-dimensional separation channel 106 and the two-dimensional separation channel 107 in the chip body 100, and solves the defect that it is difficult to fill chromatographic columns in the channels of a quartz chip in the prior art.

[0066] Specifically, in step S510, a quartz uniform glue chromium plate (quartz monolithic) coated with 500 nm thick AZ-1500 type positive photoresist and 100 nm thick chromium layer is fixed in a high-resolution contact exposure machine by standard photolithography and wet etching technology, the position of the film mask is adjusted to align with the quartz uniform glue chromium plate, and then exposure treatment is performed by ultraviolet light, so that the designed mask pattern is transferred to the quartz uniform glue chromium plate, and the uniform glue chromium plate after exposure, development and chromium removal is placed on a shaker containing hydrofluoric acid for etching, the etching depth is detected by a profilometer, and after the depth requirement is met, secondary etching is performed to prepare chip channels with different depths. The secondary etching step after glue supplementing is the same as the primary etching step, and the monolithic quartz microfluidic chip (quartz monolithic) is prepared after complete glue removal and cutting.

[0067] In step S520, the two prepared quartz plates are bonded to prepare a quartz microfluidic two-dimensional liquid chromatography chip, i.e. the chip body 100, by using plasma-assisted bonding technology and thermal bonding technology. The stacking levels of the upper and lower quartz monoliths during bonding are shown in FIG. 2. Figure 2

[0068] ​In step S530, porous silica microspheres are introduced into the connection between the plunger channel 108 and the first end of the one-dimensional separation channel 106 as a first plunger by using a tool (a fine filament with certain strength, such as a metal wire) through the plunger channel 108, and then the plunger channel 108 is sealed. A homogenate solution of chromatographic packing (such as a 3 mg / mL to 5 mg / mL C18 chromatographic packing homogenate solution) is prepared by using tetrahydrofuran as a dispersant, and ultrasonic mixing is performed until the solution is uniform. A method of air pressure driving the homogenate solution is selected to fill the one-dimensional chromatographic packed column. When the column bed length of the one-dimensional chromatographic packed column reaches a set value (such as 1 cm), the filling is stopped, and the one-dimensional chromatographic packed column is compacted at a set flow rate (such as 0.01 mL / min) and a pressure change of 0-1000 psi. After the one-dimensional chromatographic packed column is filled, a capillary bubble method is used to introduce porous silica microspheres into the connection between the two-dimensional mobile phase channel 103 and the first end of the two-dimensional separation channel 107 as a second plunger, and a two-dimensional chromatographic packed column is prepared by using the same method. Finally, a quartz microfluidic two-dimensional liquid chromatography chip containing a one-dimensional chromatographic packed column and a two-dimensional chromatographic packed column is obtained.

[0069] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application, and are not intended to limit the same. Although the present application has been described in detail with reference to the foregoing examples, those of ordinary skill in the art will understand that they can still modify the technical solutions described in the foregoing examples, or make equivalent replacements for some of the technical features thereof. Such modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A two-dimensional liquid chromatography microfluidic chip, characterized in that, The chip body is internally formed with a channel structure, and the channel structure comprises a sample channel, a one-dimensional flow phase channel, a two-dimensional flow phase channel, a one-dimensional cross-sample injection channel, a two-dimensional cross-sample injection channel, a one-dimensional separation channel, a two-dimensional separation channel, a plunger channel and a post-column channel. The sample channel is connected with a first end of the one-dimensional cross-sample injection channel, the one-dimensional flow phase channel is connected with a second end of the one-dimensional cross-sample injection channel, a first end of the one-dimensional separation channel is connected with a third end of the one-dimensional cross-sample injection channel, the plunger channel is connected with the third end of the one-dimensional cross-sample injection channel, the two-dimensional flow phase channel is connected with a first end of the two-dimensional cross-sample injection channel, a second end of the one-dimensional separation channel is connected with a second end of the two-dimensional cross-sample injection channel, a first end of the two-dimensional separation channel is connected with a third end of the two-dimensional cross-sample injection channel, and one end of the post-column channel is connected with a second end of the two-dimensional separation channel. The channel structure further comprises a first liquid discharge channel and a second liquid discharge channel, the first liquid discharge channel is connected with a fourth end of the one-dimensional cross-sample injection channel, and the second liquid discharge channel is connected with a fourth end of the two-dimensional cross-sample injection channel. A first plunger is arranged at a connection position between the plunger channel and the first end of the one-dimensional separation channel, and a second plunger is arranged at a connection position between the two-dimensional flow phase channel and the first end of the two-dimensional separation channel.

2. The two-dimensional liquid chromatography microfluidic chip of claim 1, wherein, A one-dimensional chromatographic column is arranged in the one-dimensional separation channel, and a two-dimensional chromatographic column is arranged in the two-dimensional separation channel. A first limiting channel for limiting the first plunger is arranged at an outlet of the third end of the one-dimensional cross-sample injection channel, and a second limiting channel for limiting the second plunger is arranged at an inlet of the fourth end of the two-dimensional cross-sample injection channel.

3. The two-dimensional liquid chromatography microfluidic chip of claim 2, wherein, The first limiting channel and the second limiting channel are both semicircular in cross section.

4. The two-dimensional liquid chromatography microfluidic chip of claim 3, wherein, The first limiting channel and the second limiting channel both have a diameter of 80 μm to 120 μm.

5. The two-dimensional liquid chromatography microfluidic chip of claim 4, wherein, The first plunger and the second plunger are both porous silica microspheres.

6. The two-dimensional liquid chromatography microfluidic chip of claim 2, wherein, The one-dimensional separation channel and the two-dimensional separation channel both have a diameter of 80 μm to 120 μm.

7. The two-dimensional liquid chromatography microfluidic chip according to any one of claims 1 to 6, wherein, The chip body comprises two quartz monoliths, and grooves for forming the channel structure are arranged on the two quartz monoliths, the two quartz monoliths are oppositely arranged and are bonded to each other, so that the grooves on the two quartz monoliths form the channel structure.

8. The two-dimensional liquid chromatography microfluidic chip according to any one of claims 1 to 6, wherein, The method comprises the following steps:

9. A method for fabricating a microfluidic chip based on the two-dimensional liquid chromatography microfluidic chip according to any one of claims 1 to 8, characterized in that, Grooves corresponding to the channel structure are respectively formed on two quartz monoliths; The two quartz monoliths with the grooves are aligned and heat-bonded to form the chip body, so that the grooves on the two quartz monoliths are combined to form the channel structure; and The method comprises the following steps: Grooves corresponding to the channel structure are respectively formed on two quartz monoliths; The two quartz monoliths with the grooves are aligned and heat-bonded to form the chip body, so that the grooves on the two quartz monoliths are combined to form the channel structure; and introducing a first plunger into the plunger channel through the plunger channel to the junction of the plunger channel and the first end of the one-dimensional separation channel, and then filling the one-dimensional separation channel with chromatographic packing material through the post-column channel to form a one-dimensional chromatographic packed column within the one-dimensional separation channel, introducing a second plunger into the two-dimensional mobile phase channel through the two-dimensional mobile phase channel to the junction of the two-dimensional mobile phase channel and the first end of the two-dimensional separation channel, and then filling the two-dimensional separation channel with chromatographic packing material through the post-column channel to form a two-dimensional chromatographic packed column within the two-dimensional separation channel.

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