CN116809136A - Microfluidic chip for gradient elution liquid chromatography - Google Patents
Microfluidic chip for gradient elution liquid chromatography Download PDFInfo
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- CN116809136A CN116809136A CN202310885967.1A CN202310885967A CN116809136A CN 116809136 A CN116809136 A CN 116809136A CN 202310885967 A CN202310885967 A CN 202310885967A CN 116809136 A CN116809136 A CN 116809136A
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- sample injection
- gradient elution
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- 238000010828 elution Methods 0.000 title claims abstract description 29
- 238000004811 liquid chromatography Methods 0.000 title claims abstract description 28
- 238000002347 injection Methods 0.000 claims abstract description 43
- 239000007924 injection Substances 0.000 claims abstract description 43
- 238000001514 detection method Methods 0.000 claims abstract description 37
- 238000004806 packaging method and process Methods 0.000 claims abstract description 15
- 239000000243 solution Substances 0.000 claims abstract description 11
- 238000002386 leaching Methods 0.000 claims abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 239000002699 waste material Substances 0.000 claims abstract description 6
- 239000000126 substance Substances 0.000 claims description 9
- 239000011148 porous material Substances 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 4
- 229920001169 thermoplastic Polymers 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 abstract description 15
- 238000013375 chromatographic separation Methods 0.000 abstract description 7
- 238000004587 chromatography analysis Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 239000003480 eluent Substances 0.000 description 3
- 238000010829 isocratic elution Methods 0.000 description 3
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 description 2
- 229920000089 Cyclic olefin copolymer Polymers 0.000 description 2
- 239000004713 Cyclic olefin copolymer Substances 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 102000017011 Glycated Hemoglobin A Human genes 0.000 description 1
- 108010014663 Glycated Hemoglobin A Proteins 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 230000002745 absorbent Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
Abstract
The application provides a microfluidic chip for gradient elution liquid chromatography analysis, which relates to the field of microfluidic chips and comprises the following components: an upper chip, a middle chip and a lower chip which are sequentially overlapped; the upper surface of the upper chip is sequentially provided with a leaching solution inlet, a micro-channel, a micro-mixer, a double-T-shaped sample injection channel, a sample injection hole, a sample outlet, a porous sieve plate, a micro-chromatographic column, a packaging micro-valve, a filling port, a filling channel, a Z-shaped detection tank and a waste liquid outlet; the lower surface of the upper chip is used for processing the upper layer of the micro mixer; the upper surface of the middle chip is provided with the micro-channel, the lower layer of the micro-mixer, the double-T-shaped sample injection channel, the filling channel and the micro-chromatographic column; and the Z-shaped detection pool is processed in the middle chip. The application can realize gradient elution mode liquid chromatographic separation, improve chromatographic separation efficiency and effectively reduce the sample consumption.
Description
Technical Field
The application relates to the field of microfluidic chips, in particular to a microfluidic chip for gradient elution liquid chromatography analysis.
Background
The microfluidic chip is a scientific technology with the main characteristic of controlling fluid in a micrometer scale space, and is an important development front in the fields of analysis science, micro-electromechanical processing, life science, chemical industry, analytical instruments, environmental science and the like in the new century. The liquid chromatography analysis technology is one of the important means of modern analysis, can analyze and purify molecules such as polypeptide, protein and other substances, and has important application in the fields of information science, life science, environmental science, material science and the like.
The liquid chromatography technology is transferred to the microfluidic chip, so that the analysis with small sample size and low reagent consumption can be realized, and the analysis efficiency is greatly improved. The microfluidic technology is utilized to integrate a plurality of functional units of chromatographic analysis on a chip, and has the advantages of reducing the reagent amount, shortening the analysis time and improving the detection sensitivity, and can reduce the cost and realize mass production.
When liquid chromatography is carried out, two elution modes are usually used, one is isocratic elution and the other is gradient elution. During isocratic elution analysis, the mobile phase remains unchanged throughout the process. When the gradient elution is used for chromatographic separation, the composition of mobile phases with different polarities can be continuously or intermittently changed, and the separation degree of each component in a sample is improved, so that the analysis efficiency is greatly improved.
However, microfluidic chips capable of performing gradient elution liquid chromatography have not been found on the market. In recent years, the microfluidic chip for liquid chromatography proposed in patent CN115228522a and patent CN110568087a can only perform isocratic elution. Therefore, development of a microfluidic chip capable of realizing gradient elution liquid chromatography and improvement of analysis capability are needed.
Disclosure of Invention
The application aims to provide a microfluidic chip for gradient elution liquid chromatography analysis, which is used for realizing gradient elution mode liquid chromatography separation, improving the chromatographic separation efficiency and effectively reducing the sample use amount.
In order to achieve the above object, the present application provides the following solutions:
a microfluidic chip for gradient elution liquid chromatography, comprising: an upper chip, a middle chip and a lower chip which are sequentially overlapped;
the upper surface of the upper chip is sequentially provided with a leaching solution inlet, a micro-channel, a micro-mixer, a double-T-shaped sample injection channel, a sample injection hole, a sample outlet, a porous sieve plate, a micro-chromatographic column, a packaging micro-valve, a filling port, a filling channel, a Z-shaped detection tank and a waste liquid outlet; the lower surface of the upper chip is used for processing the upper layer of the micro mixer; the upper surface of the middle chip is provided with the micro-channel, the lower layer of the micro-mixer, the double-T-shaped sample injection channel, the filling channel and the micro-chromatographic column; the Z-shaped detection pool is processed in the middle chip;
the leaching solution inlet is provided with two sections, one leaching solution inlet is connected with one section of the micro-flow channel, and the junction of the two sections of the micro-flow channels is connected with the inlet of the micro-mixer; the widths of the two sections of micro-channels are the same; the sum of the widths of the two sections of micro-channels is the same as the inlet width of the micro-mixer;
two leacheates enter from two leacheate inlets, flow through two sections of micro-channels to be converged at the inlets of the micro-mixer, and flow through the micro-mixer to obtain uniformly mixed leacheate;
the tail end of the micromixer is connected with the double-T-shaped sample injection channel, and the double-T-shaped sample injection channel is respectively connected with the sample injection hole and the sample outlet hole; the substance to be detected enters from the sample injection hole and flows out from the sample outlet through the double-T-shaped sample injection channels, and the fixed substance to be detected is filled between the double-T-shaped sample injection channels;
the two porous sieve plates are used for packaging two ends of the micro chromatographic column; the double-T-shaped sample injection channel is connected with a porous sieve plate at the front end of the micro-chromatographic column;
the front end of the micro chromatographic column is connected with the packaging micro valve filled by the micro chromatographic column, the filling channel and the filling port, the filling port is connected with the filling channel, the filling micro valve is arranged on the filling channel, and the edge of the packaging micro valve is tangential to the micro chromatographic column;
and the porous sieve plate at the rear end of the packaging micro-chromatographic column is connected with the Z-type detection pool.
Optionally, the fixed distance between the double-T-shaped sample injection channels is calculated according to chromatographic theory.
Optionally, the porous sieve plate is cylindrical and is embedded in the microfluidic chip;
the pore size of the porous sieve plate is 2 mu m.
Optionally, the aperture of the Z-shaped detection cell is 500 μm.
Optionally, a detection channel is processed in the middle chip; the detection channel is arranged in the Z-type detection pool; the detection channel is Z-shaped.
Optionally, positioning holes are machined at the same positions of the upper chip, the middle chip and the lower chip.
Optionally, the thickness of the upper chip is 5mm; the thickness of the middle chip is 3mm; the thickness of the lower chip is 2mm.
Alternatively, the microfluidic chip has a length of 70mm and a width of 30mm.
Optionally, the material of the microfluidic chip is a thermoplastic polymer.
According to the specific embodiment provided by the application, the application discloses the following technical effects: the application provides a microfluidic chip for gradient elution liquid chromatography analysis, which comprises an upper chip, a middle chip and a lower chip which are sequentially overlapped; the upper surface of the upper chip is sequentially provided with a leaching solution inlet, a micro-channel, a micro-mixer, a double-T-shaped sample injection channel, a sample injection hole, a sample outlet, a porous sieve plate, a micro-chromatographic column, a packaging micro-valve, a filling port, a filling channel, a Z-shaped detection tank and a waste liquid outlet; the lower surface of the upper chip is used for processing the upper layer of the micro mixer; the upper surface of the middle chip is provided with the micro-channel, the lower layer of the micro-mixer, the double-T-shaped sample injection channel, the filling channel and the micro-chromatographic column; and the Z-shaped detection pool is processed in the middle chip. The two leachates enter the microfluidic chip provided by the application from the two leachates inlets, so that the gradient elution mode liquid chromatographic separation can be realized, the chromatographic separation efficiency is improved, and the use amount of substances to be detected is effectively reduced.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions of the prior art, the drawings that are needed in the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Fig. 1 is a schematic top view of a microfluidic chip according to the present application;
fig. 2 is an exploded schematic view of a microfluidic chip according to the present application;
fig. 3 is a schematic front view of a microfluidic chip according to the present application;
fig. 4 is a schematic diagram of a detection flow of a microfluidic chip provided by the present application.
Detailed Description
The following description of the embodiments of the present application will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present application, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the application without making any inventive effort, are intended to be within the scope of the application.
The application aims to provide a microfluidic chip for gradient elution liquid chromatography analysis, which can realize gradient elution mode liquid chromatography separation, improve the chromatographic separation efficiency and effectively reduce the sample usage amount.
In order that the above-recited objects, features and advantages of the present application will become more readily apparent, a more particular description of the application will be rendered by reference to the appended drawings and appended detailed description.
As shown in fig. 1, the present application provides a microfluidic chip for gradient elution liquid chromatography, comprising: an upper chip, a middle chip and a lower chip which are sequentially overlapped; wherein, the same positions of the upper layer chip, the middle layer chip and the lower layer chip are processed with positioning holes 101, and three layers of chips are fixed through the positioning holes.
The upper surface of the upper chip is sequentially provided with a leaching solution inlet 102, a micro-channel 103, a micro-mixer 104, a double-T-shaped sample injection channel 106, a sample injection hole 105, a sample outlet 107, a porous sieve plate 108, a micro-chromatographic column 109, a packaging micro-valve 110, a filling channel 111, a filling port 112, a Z-shaped detection pool 113 and a waste liquid outlet; the lower surface of the upper chip processes the upper layer of the micromixer 104; the upper surface of the middle chip is provided with the micro flow channel 103, the lower layer of the micro mixer 104, the double-T-shaped sample injection channel 106, the filling channel 111 and the micro chromatographic column 109; the Z-shaped detection cell 113 is fabricated in the middle chip.
The eluent inlet 102 is connected with the micro-flow channel 103, the two sections of micro-flow channels 103 are connected with the inlet of the micro-mixer 104 after being converged, the two sections of micro-flow channels 103 have the same width, and the sum of the widths is the same as the width of the inlet of the micro-mixer 104; the two leacheates enter from two inlets of the leacheate, flow through the micro-channel 103 and are converged at the inlet of the micro-mixer 104, and the mixed leacheate is obtained after flowing through the micro-mixer 104.
The end of the micromixer 104 is connected with a double-T-shaped sample injection channel 106, the double-T-shaped sample injection channel 106 is respectively connected with a sample injection hole 105 and a sample outlet hole 107, and the fixed distance between the double-T-shaped sample injection channels 106 is calculated according to chromatographic theory. The substance to be detected enters from the sample injection hole 105 and flows out from the sample outlet 107 through the double T-shaped sample injection channels 106, and the fixed substance to be detected is filled between the double T-shaped sample injection channels 106.
The tail end of the double T-shaped sample injection channel 106 is connected with a porous sieve plate 108, the two porous sieve plates 108 are used for packaging the two ends of the micro-chromatographic column 109, the porous sieve plate 108 is embedded in a chip in a cylindrical shape, the porous sieve plate 108 can bear tens of megapascals of pressure, the pore is 2 mu m, and the small-pore porous sieve plate 108 can package chromatographic packing microspheres with the diameter larger than 2 mu m.
The front end of the micro chromatographic column 109 is connected with a packed micro valve 110 filled with chromatographic column, a filling channel 111 and a filling port 112, the filling port 112 is connected with the filling channel 111, the filling channel 111 is provided with a packed micro valve, and the edge of the packed micro valve 110 is tangential to the micro chromatographic column 109.
The porous sieve plate 108 of the packed micro-chromatographic column 109 is connected with a Z-shaped detection cell 113, the aperture size of the Z-shaped detection cell 113 is 500 mu m, and the Z-shaped detection cell 113 is processed on a micro-fluidic chip similarly to the Z-shaped detection cell 113 of a commercial chromatograph, so that the efficiency of chromatographic analysis is improved.
The length of the microfluidic chip is 70mm, and the width of the microfluidic chip is 30mm.
As shown in fig. 2-3, the upper chip is processed with an eluent inlet 102, a sample inlet 105, a sample outlet 107, a filling port 112 and a packaging micro valve 110; the lower surface of the upper chip is also provided with a micromixer upper layer 301; the upper surface of the middle chip is provided with a micro-channel 103, a micro-mixer lower layer 302, a double-T-shaped sample injection channel 106, a filling channel 111 and a micro-chromatographic column 109; a Z-type detection cell 113 is further processed on a middle chip, and a detection channel 303 is processed in the middle chip; the detection channel 303 is arranged in the Z-type detection cell 113; the detection channel 303 is Z-shaped.
The thickness of the upper chip is 5mm, the thickness of the middle chip is 3mm, and the thickness of the lower chip is 2mm.
The material of the microfluidic chip provided by the application is a thermoplastic polymer.
As a specific example of the present application, the thermoplastic polymer material is a Cyclic Olefin Copolymer (COC).
After the microstructure on the three-layer chip is processed, the three-layer chip is bonded by adopting a laser fusion bonding process under the irradiation of 9.5KW power laser after the light absorbent is smeared on the surface of the chip.
Fig. 4 is a schematic diagram of a detection flow of a microfluidic chip provided by the present application, as shown in fig. 4, two injection pumps 401 pump two kinds of leacheate from the leacheate inlet 102 of the microfluidic chip 404 provided by the present application (different flow rate ratios may be set 1:9, 2:8, 3:7 and … …). The two rinse streams are thoroughly mixed after passing through the micromixer 104. The two syringe pumps 401 are turned off and a syringe 402 is used to inject the sample from the inlet of the dual tee injection channel 106. The syringe pump 401 is turned on and the well mixed eluent pushes the immobilized trapped sample into the micro-chromatography column 109. After separation by a chromatographic column, the detected component flows through a Z-shaped detection cell 113, at which point the sample component is detected by an absorbance detector, and the result is obtained by computer data analysis. The application also includes: sample waste collection vial 403, light source 405, detector 406, and data processor 407.
The chip can be used for detecting the proportions of HbA, hbA2 and HbF of the components of hemoglobin (Hb) under 415nm light, and can be used for accurately detecting the HbA1c component of glycosylated hemoglobin. The chip is used for accurately detecting the content of bilirubin under 445nm illumination.
The gradient elution liquid chromatography micro-fluidic chip can realize absorbance detection.
The detection wavelength range is 320-700nm.
The sample injection amount was 0.62. Mu.L.
The application realizes the gradient elution mode liquid chromatography analysis on the micro-fluidic chip 404, can effectively improve the chromatography efficiency and reduce the sample usage amount.
In the present specification, each embodiment is described in a progressive manner, and each embodiment is mainly described in a different point from other embodiments, and identical and similar parts between the embodiments are all enough to refer to each other.
The principles and embodiments of the present application have been described herein with reference to specific examples, the description of which is intended only to assist in understanding the methods of the present application and the core ideas thereof; also, it is within the scope of the present application to be modified by those of ordinary skill in the art in light of the present teachings. In view of the foregoing, this description should not be construed as limiting the application.
Claims (9)
1. A microfluidic chip for gradient elution liquid chromatography, comprising: an upper chip, a middle chip and a lower chip which are sequentially overlapped;
the upper surface of the upper chip is sequentially provided with a leaching solution inlet, a micro-channel, a micro-mixer, a double-T-shaped sample injection channel, a sample injection hole, a sample outlet, a porous sieve plate, a micro-chromatographic column, a packaging micro-valve, a filling port, a filling channel, a Z-shaped detection tank and a waste liquid outlet; the lower surface of the upper chip is used for processing the upper layer of the micro mixer; the upper surface of the middle chip is provided with the micro-channel, the lower layer of the micro-mixer, the double-T-shaped sample injection channel, the filling channel and the micro-chromatographic column; the Z-shaped detection pool is processed in the middle chip;
the leaching solution inlets are two, the micro-flow channel is provided with two sections, one leaching solution inlet is connected with one section of micro-flow channel, and the junction of the two sections of micro-flow channels is connected with the inlet of the micro-mixer; the widths of the two sections of micro-channels are the same; the sum of the widths of the two sections of micro-channels is the same as the inlet width of the micro-mixer;
two leacheates enter from two leacheate inlets, flow through two sections of micro-channels to be converged at the inlets of the micro-mixer, and flow through the micro-mixer to obtain uniformly mixed leacheate;
the tail end of the micromixer is connected with the double-T-shaped sample injection channel, and the double-T-shaped sample injection channel is respectively connected with the sample injection hole and the sample outlet hole; the substance to be detected enters from the sample injection hole and flows out from the sample outlet through the double-T-shaped sample injection channels, and the fixed substance to be detected is filled between the double-T-shaped sample injection channels;
the two porous sieve plates are used for packaging two ends of the micro chromatographic column; the double-T-shaped sample injection channel is connected with a porous sieve plate at the front end of the micro-chromatographic column;
the front end of the micro chromatographic column is connected with the packaging micro valve filled by the micro chromatographic column, the filling channel and the filling port, the filling port is connected with the filling channel, the filling micro valve is arranged on the filling channel, and the edge of the packaging micro valve is tangential to the micro chromatographic column;
and the porous sieve plate at the rear end of the packaging micro-chromatographic column is connected with the Z-type detection pool.
2. The microfluidic chip for gradient elution liquid chromatography according to claim 1, wherein the fixed distance between the double-T-shaped sample injection channels is calculated according to chromatographic theory.
3. The microfluidic chip for gradient elution liquid chromatography according to claim 1, wherein said porous sieve plate is cylindrical and is embedded in the microfluidic chip;
the pore size of the porous sieve plate is 2 mu m.
4. The microfluidic chip for gradient elution liquid chromatography according to claim 1, wherein the pore size of the Z-shaped detection cell is 500 μm.
5. The microfluidic chip for gradient elution liquid chromatography according to claim 1, wherein a detection channel is processed in the middle chip; the detection channel is arranged in the Z-type detection pool; the detection channel is Z-shaped.
6. The microfluidic chip for gradient elution liquid chromatography according to claim 1, wherein positioning holes are processed at the same positions of the upper chip, the middle chip and the lower chip.
7. The microfluidic chip for gradient elution liquid chromatography according to claim 1, wherein the thickness of said upper chip is 5mm; the thickness of the middle chip is 3mm; the thickness of the lower chip is 2mm.
8. The microfluidic chip for gradient elution liquid chromatography according to claim 1, wherein the length of the microfluidic chip is 70mm and the width is 30mm.
9. The microfluidic chip for gradient elution liquid chromatography according to claim 1, wherein the material of the microfluidic chip is a thermoplastic polymer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310885967.1A CN116809136A (en) | 2023-07-19 | 2023-07-19 | Microfluidic chip for gradient elution liquid chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310885967.1A CN116809136A (en) | 2023-07-19 | 2023-07-19 | Microfluidic chip for gradient elution liquid chromatography |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116809136A true CN116809136A (en) | 2023-09-29 |
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ID=88118406
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310885967.1A Pending CN116809136A (en) | 2023-07-19 | 2023-07-19 | Microfluidic chip for gradient elution liquid chromatography |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116809136A (en) |
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2023
- 2023-07-19 CN CN202310885967.1A patent/CN116809136A/en active Pending
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