CN110568087A - high-sensitivity detection device and method for diabetes marker - Google Patents
high-sensitivity detection device and method for diabetes marker Download PDFInfo
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Abstract
The invention provides a high-sensitivity detection device and method for a diabetes marker. The high-pressure liquid chromatography pump provides driving pressure to pump the buffer solution into a pipeline of the droplet microfluidic chip; introducing the HbA1c sample from the sample inlet through the sample injection valve to finish quantitative injection; separating each component in the HbA1c sample by using a strong cation exchange polymer monolithic column of the droplet microfluidic chip; the droplet generator wraps the separated HbA1c component to form a water-in-oil droplet and flows into the micro detection cell; the deuterium tungsten-halide lamp light source provides ultraviolet light; the ultraviolet spectrometer receives ultraviolet light to detect the absorbance of the HbA1c sample, and transmits a detection signal to data processing software of a computer through an optical fiber to perform data processing to obtain the concentration ratio of the HbA1 c. The method improves the detection sensitivity of HbA1c, is simple to operate, and is beneficial to on-site real-time detection.
Description
Technical Field
The invention relates to the technical field of diabetes marker detection, in particular to a high-sensitivity detection device and method for a diabetes marker.
Background
Diabetes is a group of metabolic diseases characterized by hyperglycemia, has high morbidity, and is one of four main diseases which harm human health at present. Diabetic ketosis, cardiovascular diseases, neuropathy, diabetic foot, diabetic nephropathy and other complications are easily caused by long-term diabetes, and the health of human beings is seriously threatened. According to the publication of the diabetes society of the Chinese medical society in 2013, 1.39 hundred million diabetic patients are estimated in China, and 60.7 percent of the diabetic patients are not diagnosed.
For many years, diabetes diagnosis has been based on plasma glucose levels, including fasting glucose and 2-hour blood glucose values from the oral glucose tolerance test. However, the blood sugar test method is susceptible to diet, exercise, drugs, and other factors and requires a specific time for blood collection, which greatly limits the accuracy of test results and also brings inconvenience to patients. The detection methods of glycated hemoglobin (HbA1c) applied in clinical laboratories at present mainly have two main categories: one class of methods is based on the difference in charge between glycated hemoglobin and non-glycated hemoglobin, such as ion exchange chromatography, electrophoresis, and the like; another class of methods is based on the structural features of the glycated groups on hemoglobin, such as affinity chromatography, ion trapping, and immunization. In 2002, the international association of clinical chemistry (IFCC) published the international standard method for HbA1c detection, i.e. a combined detection method by high pressure liquid chromatography and mass spectrometry (HPLC-MS). However, the HPLC-MS combined detection method requires expensive instruments and complicated operations, making it difficult to be clinically generalized. Currently, ion exchange HPLC is considered to be a gold standard for analytical detection of HbA1c, which measures the percentage of HbA1c in the blood relative to total hemoglobin (Hb). However, this method employs a commercial cation exchange chromatography column in combination with an external detection method, resulting in a large reagent consumption and a long analysis time, and a large dead volume exists between the end of the chromatography column and the detector due to the pipe connection, so that the separated sample components are easily diffused and remixed, thereby decreasing the detection sensitivity. Therefore, the prior art has the defects of low detection sensitivity, large detection instrument volume, complex operation steps and the like.
disclosure of Invention
the invention mainly aims to provide a device and a method for detecting a diabetes marker with high sensitivity, and aims to solve the technical problem that the prior art has low detection sensitivity on the diabetes marker.
In order to achieve the above object, the present invention provides a diabetes marker high-sensitivity detection device, comprising a droplet microfluidic chip, a high-pressure liquid chromatography pump, a sample injection valve, a deuterium halogen tungsten lamp light source, an ultraviolet spectrometer and a computer, wherein:
The high-pressure liquid chromatography pump is used for providing driving pressure to pump the buffer solution into a pipeline of the droplet microfluidic chip; the sample injection valve is used for introducing the HbA1c sample from the sample inlet and enabling the HbA1c sample to flow out from the sample waste liquid outlet, so that quantitative sample injection is completed; the droplet microfluidic chip comprises a strong cation exchange polymer monolithic column, a droplet generator and a micro detection pool; the strong cation exchange polymer monolith is used for separating each component in the HbA1c sample; the droplet generator is used for wrapping the separated HbA1c component to form a series of water-in-oil droplets with different concentrations; the micro detection cell is aligned with the light path of the ultraviolet spectrometer; the deuterium tungsten halide lamp light source is used for providing ultraviolet light for detecting the absorbance of the HbA1c sample; the ultraviolet spectrometer is used for receiving ultraviolet light to carry out absorbance detection on the HbA1c sample and transmitting a detection signal to a computer through an optical fiber; and the computer is provided with data processing software for carrying out data processing on the detection signals to obtain the HbA1c concentration ratio.
Preferably, the droplet microfluidic chip is composed of a cover plate layer, an upper fluid channel layer, a lower fluid channel layer and a substrate layer in sequence.
preferably, the cover sheet layer is provided with a water phase inlet, a first oil phase inlet, a second oil phase inlet and a fluid outlet, and the water phase inlet penetrates through the cover sheet layer and is communicated with the water phase introducing channel on the upper fluid channel layer.
Preferably, the shape of the aqueous phase introduction channel is a serpentine shape, and the interior of the aqueous phase introduction channel is filled with a strong cation exchange polymer monolithic column.
Preferably, the aqueous phase introduction channel is internally provided with a chromatographic column, and the first oil phase inlet and the second oil phase inlet penetrating through the cover plate layer respectively pass through the first oil phase introduction channel and the second oil phase introduction channel on the upper fluid channel layer and form a cross-shaped channel with the aqueous phase introduction channel and the first droplet channel.
Preferably, the cross-shaped channel is a constricted structure with an arc shape, and the width of the cross-shaped channel satisfies the relations of 2 × W1 ≧ W2 ≧ 0.5 × W1, 2 × W1 ≧ W3 ≧ W4 ≧ 0.5 × W1, and W1> W2, where × denotes a multiplication number, W1 is the terminal channel width of the aqueous phase introduction channel, W2 is the inlet channel width of the first droplet channel, W3 is the terminal channel width of the first oil phase introduction channel, W4 is the terminal channel width of the second oil phase introduction channel, and W is the front channel width of the first droplet channel.
Preferably, the upper fluid channel layer is provided with a first small hole, the lower fluid channel layer is provided with a second small hole, the first small hole penetrating through the upper fluid channel layer corresponds to the second small hole penetrating through the lower fluid channel layer in position, and the first small hole and the second small hole penetrate through the upper fluid channel layer to form the micro detection cell.
Preferably, the micro detection cell is communicated with a first droplet channel on the upper fluid channel layer and a second droplet channel penetrating through the lower fluid channel layer, and the second droplet channel is communicated with the outside through a fluid outlet sequentially penetrating through the lower fluid channel layer, the upper fluid channel layer and the cover sheet layer.
Preferably, the droplet microfluidic chip is made of one or more of polymethyl methacrylate (PMMA), Polycarbonate (PC), Polystyrene (PS) and Cyclic Olefin Copolymer (COC).
In another aspect, the present invention also provides a method for detecting a diabetes marker using the diabetes marker high-sensitivity detection apparatus, the method comprising the steps of: the high-pressure liquid chromatography pump provides a driving force to control the fluid, and a buffer solution carrying the sample is pumped into the droplet microfluidic chip from the sample injection valve so that the buffer solution drives the HbA1c sample to complete the separation of each component; wrapping the separated HbA1c sample components with an oil phase to create water-in-oil droplets; enabling the water-in-oil droplets to flow into a miniature detection pool, and detecting HbA1c sample components in the water-in-oil droplets in the miniature detection pool by using an ultraviolet spectrometer; carrying out absorbance analysis on each component wrapped in the water-in-oil droplets by using an ultraviolet spectrometer to obtain a chromatogram of the separated component; the HbA1c concentration ratio is obtained by calculating the peak area of HbA1c in the chromatogram to the peak area of total Hb by using data processing software of a computer.
compared with the prior art, the device and the method for detecting the diabetes marker with high sensitivity adopt the technical scheme, and the following technical effects are achieved: the liquid drop micro-fluidic chip for the high-sensitivity detection device of the diabetes marker has the advantages of small volume, low cost and simple operation, and is beneficial to on-site real-time detection. According to the invention, the cation exchange chromatographic column and the detection pool are integrated on the microfluidic chip, and the droplet generator is additionally arranged between the tail end of the chromatographic column and the detection pool, so that not only is the dead volume generated by pipeline connection avoided, but also the sample is prevented from being diffused and remixed at the tail end of the chromatographic column by means of droplet detection, and thus the detection sensitivity is improved. In addition, the structure of the miniature detection cell is designed to be Z-shaped, so that the optical path is increased, and the detection sensitivity is further improved. The liquid drop microfluidic chip is not only suitable for high-sensitivity detection of HbA1c, but also generally suitable for detection of biomacromolecules such as polypeptide, nucleic acid and protein with absorption peaks in ultraviolet light wave bands.
drawings
FIG. 1 is a schematic structural diagram of a high-sensitivity detection device for diabetes markers according to the present invention;
FIG. 2 is a schematic structural diagram of a droplet microfluidic chip of the high-sensitivity detection device for diabetes markers of the present invention;
FIG. 3 is a schematic cross-sectional view of a droplet microfluidic chip of the high-sensitivity detection device for diabetes markers of the present invention;
FIG. 4 is a schematic structural diagram of a droplet generator of a droplet microfluidic chip of the high-sensitivity detection device for diabetes markers of the present invention;
FIG. 5 is a flow chart of the detection method of the diabetes mellitus high-sensitivity detection device of the invention.
the implementation, functional features and advantages of the objects of the present invention will be further explained with reference to the accompanying drawings.
Detailed Description
To further explain the technical means and effects of the present invention adopted to achieve the predetermined objects, the following detailed description of the embodiments, structures, features and effects of the present invention will be given with reference to the accompanying drawings and preferred embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
referring to fig. 1, fig. 1 is a schematic structural diagram of a diabetes marker high-sensitivity detection device according to the present invention. In this embodiment, the diabetes marker high-sensitivity detection device includes a droplet microfluidic chip 1, a high-pressure liquid chromatography pump 2, a sample injection valve 3, a deuterium tungsten halogen lamp light source 10, an ultraviolet spectrometer 11 and a computer 13; the high-pressure liquid chromatography pump 2 provides driving pressure to pump the buffer solution 6 into the pipeline of the droplet microfluidic chip 1; the sample injection valve 3 is used for introducing a glycosylated hemoglobin (HbA1c) sample from a sample inlet 4 and discharging the glycosylated hemoglobin from a sample waste liquid outlet 5 to finish quantitative sample injection; the droplet microfluidic chip 1 comprises a strong cation exchange polymer monolithic column 7, a droplet generator 8 and a micro detection cell 9; the strong cation exchange polymer monolith 7 was used to separate the components in the HbA1c sample; the droplet generator 8 is used for wrapping the separated HbA1c component to form a series of water-in-oil droplets with different concentrations; the micro detection cell 9 is aligned with the light path of the ultraviolet spectrometer 11; the deuterium tungsten halide lamp light source 10 provides ultraviolet light for detecting the absorbance of the HbA1c sample; the ultraviolet spectrometer 11 is used for receiving ultraviolet light to perform absorbance detection on the HbA1c sample and transmitting a detection signal to the computer 13 through the optical fiber 12; the computer 13 is provided with data processing software for processing the detection signal to obtain the HbA1c concentration ratio.
Referring to fig. 2, fig. 2 is a schematic structural view of a droplet microfluidic chip of the diabetes marker high-sensitivity detection device of the present invention. In this embodiment, the droplet microfluidic chip 1 is composed of a cover sheet layer 20, an upper fluid channel layer 30, a lower fluid channel layer 40, and a substrate layer 50 in sequence; the cover sheet layer 20 is provided with a water phase inlet 21, a first oil phase inlet 22, a second oil phase inlet 23 and a fluid outlet 24. The water phase inlet 21 penetrates through the cover sheet layer 20 and is communicated with a water phase introducing channel 31 on the upper fluid channel layer 30; a chromatographic column is arranged inside the water phase introducing channel 31; the first oil phase inlet 22 and the second oil phase inlet 23 penetrating the cover sheet layer 20 pass through the first oil phase introduction passage 32 and the second oil phase introduction passage 33 on the upper fluid passage layer 30, respectively, and the droplet generator 8 forming a cross-shaped passage with the aqueous phase introduction passage 31 and the first droplet passage 34, and the droplet generator 8 is formed by the first oil phase introduction passage 32 and the second oil phase introduction passage 33, and the cross-shaped passage with the aqueous phase introduction passage 31 and the first droplet passage 34.
As a preferable mode, the shape of the aqueous phase introduction channel 31 is a serpentine shape, and a strong cation exchange polymer monolithic column is filled in the aqueous phase introduction channel, the strong cation exchange polymer monolithic column is a chromatographic column, the material of the strong cation exchange polymer monolithic column is acrylate copolymer, and the interior of the strong cation exchange polymer monolithic column is loose and porous; the strong cation exchange polymer monolithic column is generated by light or heat initiated polymerization reaction, and the performance parameters of the monolithic column, such as mechanical strength, pore size, pH value tolerance range, exchange capacity and the like, can be adjusted by changing the type and the proportion of the polymer. The monolith channel length may be set in the range of 1 to 25 cm, the channel depth may be set in the range of 0.05 mm to 1mm, and the channel width may be set in the range of 0.05 mm to 2 mm. The invention provides a chromatographic separation column with loose and porous surface, low back pressure, large specific surface area, high column capacity and good permeability, and the column parameters of the chromatographic column, such as the pore size and the stationary phase particle size, can be controlled by controlling the type, proportion and initiation conditions of prepolymer, thereby realizing the separation effect of different functions. The main functional components of the chip of the strong cation exchange polymer monolithic column are composed of the polymer monolithic column, the droplet generator 8 and the micro detection pool 9, and the size and the pore structure of the polymer monolithic column, the width of the droplet generator 8 and the effective length of the micro detection pool 9 can be changed according to different requirements in practical application.
Referring to fig. 3, fig. 3 is a schematic cross-sectional structure view of a droplet microfluidic chip of the diabetes marker high-sensitivity detection device of the present invention. In this embodiment, the first small hole 35 penetrating the upper fluid channel layer 30 and the second small hole 41 penetrating the lower fluid channel layer 40 are positioned correspondingly, and both are penetrated to form the micro detection cell 9, and the structure of the micro detection cell 9 is designed to be Z-shaped. The micro detection cells 9 communicate with the first droplet channels 34 on the upper fluid channel layer 30 and with the second droplet channels 42 that extend through the lower fluid channel layer 40; the second droplet channels 42 communicate with the outside through the fluid outlets 24 which pass through the lower fluid channel layer 40, the upper fluid channel layer 30 and the cover sheet layer 20 in this order.
Referring to fig. 4, fig. 4 is a schematic structural diagram of a droplet generator of a droplet microfluidic chip of the diabetes marker high-sensitivity detection device of the present invention. Preferably, the droplet generator 8 is a cross-shaped passage formed by the first oil phase introduction passage 32 and the second oil phase introduction passage 33, and the aqueous phase introduction passage 31 and the first droplet passage 34. The cross-shaped channel is of an arc-shaped contraction structure, so that small-size liquid drops can be generated, and the width of the cross-shaped channel meets the relations that 2W 1 is more than or equal to W2 is more than or equal to 0.5W 1, 2W 1 is more than or equal to W3, W4 is more than or equal to 0.5W 1, and W1 is more than W2; where denotes a multiplication number, W1 is the terminal channel width of the aqueous phase introduction channel 31, W2 is the inlet channel width of the first droplet channel 34, W3 is the terminal channel width of the first oil phase introduction channel 32, W4 is the terminal channel width of the second oil phase introduction channel 33, and W is the width of the first droplet channel 34; the working principle is as follows: the oil phase and the aqueous phase solutions are separately injected from the respective inlets using an external pump or a valve driving system, and at the cross-channel intersection, water-in-oil (W/O) droplets are generated due to the interaction between the forces such as shear force, viscous force, surface tension, and inertial force, and are carried into the first droplet channel 34 by the oil phase, as the aqueous phase is squeezed by the oil phase. The size of the droplets generated by the droplet generator 8 can be changed by adjusting the flow rate ratio of the fluid in the first oil phase introduction passage 32 and the second oil phase introduction passage 33 to the water phase introduction passage 31.
in this embodiment, the droplet microfluidic chip 1 may be made of a transparent thermoplastic material, such as: one or more of polymethyl methacrylate (PMMA), Polycarbonate (PC), Polystyrene (PS) and Cyclic Olefin Copolymer (COC). The structure and size of the droplet microfluidic chip 1 made of different materials are described in the following specific examples.
as a first preferred embodiment, the material of the droplet microfluidic chip 1 is PMMA, and the droplet microfluidic chip 1 includes four layers of simple planar structures: cover sheet layer 20, upper fluid channel layer 30, lower fluid channel layer 40, and base layer 50. The thickness of the cover layer 20 and the thickness of the substrate layer 50 are both 2mm, the thickness of the upper fluid channel layer 30 and the thickness of the lower fluid channel layer 40 are both 1mm, and the length of the polymer integral column is about 50mm, the width is 0.5mm, and the height is 0.5 mm; the cross channel of the droplet generator 8 is 0.5mm high, W1W 3W 4W 0.2mm, W0.5 mm; the diameter of the micro detection pool 9 is 1mm, and the height thereof is 2 mm; the water phase inlet 21, first oil phase inlet 22, second oil phase inlet 23 and fluid outlet 24 are all 1.6mm in diameter.
As a second preferred embodiment, the material of the droplet microfluidic chip 1 is COC, the thicknesses of the cover sheet layer 20 and the substrate layer 50 of the droplet microfluidic chip 1 are both 2mm, the thicknesses of the upper fluid channel layer 30 and the lower fluid channel layer 40 are both 0.5mm, the length of the polymer monolithic column is about 30mm, the width is 0.5mm, and the height is 0.3 mm; the cross channel of the droplet generator 8 is 0.3mm high, W1W 3W 4W 363 mm, W0.5 mm; the diameter of the micro detection pool 9 is 0.6mm, and the height is 1 mm; the water phase inlet 21, first oil phase inlet 22, second oil phase inlet 23 and fluid outlet 24 are all 1.0mm in diameter.
In the droplet microfluidic chip 1 of the present invention, the buffer solution loaded with the sample enters the bent serpentine aqueous phase introduction channel 31 from the aqueous phase inlet 11, and the bent serpentine structure can increase the length of the polymer monolithic column in the aqueous phase introduction channel 31 without increasing the length of the droplet microfluidic chip, thereby facilitating high-throughput separation and analysis of the sample. Moreover, the polymer monolithic column has the advantages of wide pH value tolerance range, small pressure drop, porosity, high exchange capacity and the like, the requirement on the high output pressure of a driving device is greatly reduced, and more choices are provided for a mobile phase. In addition, the polymer monolithic columns with different pore diameters and specific surface areas can be prepared by adjusting the proportion of the prepolymer, which is beneficial to improving the separation efficiency and the separation speed. After the sample is subjected to chromatographic separation in the polymer monolithic column, the sample components flowing out of the tail end of the chromatographic column are wrapped in the form of liquid drops by the oil solutions injected from the first oil phase introduction channel 32 and the second oil phase introduction channel 33 under the action of shearing force, surface tension, viscous force and inertia force, so that the separated components are prevented from molecular diffusion before entering the micro detection cell 9, the peak band broadening effect and the remixing of the separated components are avoided, and the separation effect is improved. In addition, the micro detection cell 9 is integrated on the droplet microfluidic chip, so that the absorbance of the separated components can be monitored in real time and on line, the real-time concentration of the sample can be obtained, the micro detection cell 9 penetrates through the upper fluid channel layer 30 and the lower fluid channel layer 40, the optical path of the micro detection cell 9 is increased, the full-transparent detection cell is realized, and the detection sensitivity is greatly improved.
On the other hand, the invention also provides a detection method based on the diabetes high-sensitivity detection device. Referring to fig. 5, fig. 5 is a flow chart of a preferred embodiment of the detecting method of the diabetes mellitus high-sensitivity detecting device of the invention, which comprises the following steps:
and step S51, the high-pressure liquid chromatography pump 2 provides a driving force to control the fluid, and the buffer solution carrying the sample is pumped into the droplet microfluidic chip 1 from the sample injection valve 3, so that the buffer solution drives the HbA1c sample to complete the separation of each component. In this embodiment, the fluid is controlled by an external pump or valve to provide a driving force to pump the sample-carrying buffer solution from the aqueous phase inlet 21 into the aqueous phase introduction channel 31, and the separated components sequentially flow out of the polymer monolith due to the difference in the moving speed of the sample in the polymer monolith caused by the difference in the forces of the different components in the sample and the polymer monolith in the aqueous phase introduction channel 31.
At step S52, the separated HbA1c sample components are encapsulated by an oil phase to produce water-in-oil droplets. In this embodiment, the oil solution is merged with the component aqueous solution separated from the end of the aqueous phase introduction passage 31 at the cross passage from the first oil phase inlet 22 and the second oil phase inlet 23 through the first oil phase introduction passage 32 and the second oil phase introduction passage 33, respectively, and water-in-oil droplets with controllable particle diameters are formed by controlling the flow rate ratio of the oil solution and the aqueous solution to enter the first droplet passage 34.
Step S53, enabling the water-in-oil droplets to flow into a micro detection pool 9, and detecting HbA1c sample components in the water-in-oil droplets in the micro detection pool 9 by using an ultraviolet spectrometer 11; finally, the water-in-oil droplet subjected to absorbance detection flows out of the fluid outlet 24 via the second droplet channel 42.
step S54, performing absorbance analysis on each component wrapped in the water-in-oil droplets to obtain an ion chromatogram based on droplet detection, namely a chromatogram of the separated component. The ultraviolet spectrometer 11 is configured to receive ultraviolet light, transmit a signal to data processing software of the computer 13 through the optical fiber 12, perform data processing, finally complete absorbance detection, and perform absorbance analysis on each component encapsulated in the water-in-oil droplet, thereby obtaining an ion chromatogram based on droplet detection.
Step S55, the HbA1c concentration ratio is obtained by calculating the peak area of HbA1c in the chromatogram to the peak area of total Hb. In the present embodiment, the data processing software of the computer 13 obtains the HbA1c concentration ratio by calculating the peak area of HbA1c to the peak area of total Hb (hemoglobin) in the chromatogram.
The liquid drop micro-fluidic chip in the high-sensitivity detection device for the diabetes marker provided by the invention has the advantages of small volume, low cost and simple operation, and is beneficial to on-site real-time detection. In addition, the invention also provides a high-sensitivity detection method for the diabetes marker, which is not only suitable for high-sensitivity detection of the HbA1c sample, but also suitable for detection of biological macromolecules such as polypeptide, nucleic acid and protein. In order to improve the detection sensitivity, the cation exchange chromatographic column and the detection pool are integrated on the microfluidic chip, and the droplet generator is additionally arranged between the tail end of the chromatographic column and the detection pool. In addition, the structure of the miniature detection cell is designed to be Z-shaped, so that the optical path is increased, and the detection sensitivity is further improved. The liquid drop microfluidic chip is not only suitable for high-sensitivity detection of HbA1c, but also generally suitable for detection of biomacromolecules such as polypeptide, nucleic acid and protein with absorption peaks in ultraviolet light wave bands.
The above description is only a preferred embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by using the contents of the present specification and the accompanying drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (10)
1. The utility model provides a diabetes marker high sensitivity detection device, its characterized in that, this diabetes marker high sensitivity detection device includes liquid drop micro-fluidic chip, high-pressure liquid chromatography pump, injection valve, deuterium halogen tungsten lamp light source, ultraviolet ray spectrum appearance and computer, wherein:
the high-pressure liquid chromatography pump is used for providing driving pressure to pump the buffer solution into a pipeline of the droplet microfluidic chip;
The sample injection valve is used for introducing the HbA1c sample from the sample inlet and enabling the HbA1c sample to flow out from the sample waste liquid outlet, so that quantitative sample injection is completed;
The droplet microfluidic chip comprises a strong cation exchange polymer monolithic column, a droplet generator and a micro detection pool; the strong cation exchange polymer monolith is used for separating each component in the HbA1c sample; the droplet generator is used for wrapping the separated HbA1c component to form a series of water-in-oil droplets with different concentrations; the micro detection cell is aligned with the light path of the ultraviolet spectrometer;
The deuterium tungsten halide lamp light source is used for providing ultraviolet light for detecting the absorbance of the HbA1c sample;
The ultraviolet spectrometer is used for receiving ultraviolet light to carry out absorbance detection on the HbA1c sample and transmitting a detection signal to a computer through an optical fiber;
And the computer is provided with data processing software for carrying out data processing on the detection signals to obtain the HbA1c concentration ratio.
2. The diabetes marker high-sensitivity detection device according to claim 1, wherein the droplet microfluidic chip is composed of a cover plate layer, an upper fluid channel layer, a lower fluid channel layer and a substrate layer in sequence.
3. The diabetes marker high-sensitivity detection device according to claim 2, wherein the cover sheet layer is provided with a water phase inlet, a first oil phase inlet, a second oil phase inlet and a fluid outlet, and the water phase inlet penetrates through the cover sheet layer and is communicated with the water phase introduction channel on the upper fluid channel layer.
4. The diabetes marker high-sensitivity detection device according to claim 3, wherein the aqueous phase introduction channel is in a serpentine shape, and a strong cation exchange polymer monolithic column is filled in the aqueous phase introduction channel.
5. the diabetes marker high-sensitivity detection device according to claim 3, wherein the aqueous phase introduction channel is internally provided with a chromatographic column, and the first oil phase inlet and the second oil phase inlet penetrating through the cover sheet layer respectively pass through the first oil phase introduction channel and the second oil phase introduction channel on the upper fluid channel layer and form a cross-shaped channel with the aqueous phase introduction channel and the first droplet channel.
6. The diabetes marker high-sensitivity detection device according to claim 3, wherein the cross-shaped channel has an arc-shaped constriction structure, and the width of the cross-shaped channel satisfies the relationship of 2W 1 ≧ W2 ≧ 0.5W 1, 2W 1 ≧ W3 ≧ W4 ≧ 0.5W 1, and W1> W2, where × denotes a multiplication number, W1 is the terminal channel width of the aqueous phase introduction channel, W2 is the inlet channel width of the first droplet channel, W3 is the terminal channel width of the first oil phase introduction channel, W4 is the terminal channel width of the second oil phase introduction channel, and W is the front end channel width of the first droplet channel.
7. The diabetes marker high-sensitivity detection device according to claim 3, wherein the upper fluid channel layer is provided with a first small hole, the lower fluid channel layer is provided with a second small hole, the first small hole penetrating through the upper fluid channel layer corresponds to the second small hole penetrating through the lower fluid channel layer in position, and the first small hole and the second small hole form the micro detection pool after being penetrated.
8. The diabetes marker high-sensitivity detection device according to claim 7, wherein the micro detection cell is communicated with a first droplet channel on the upper fluid channel layer and a second droplet channel penetrating through the lower fluid channel layer, and the second droplet channel is communicated with the outside through a fluid outlet which sequentially penetrates through the lower fluid channel layer, the upper fluid channel layer and the cover sheet layer.
9. The diabetes marker high-sensitivity detection device according to any one of claims 1 to 8, wherein the droplet microfluidic chip is made of one or more of polymethyl methacrylate, polycarbonate, polystyrene and cyclic olefin copolymer.
10. A method for detecting a diabetes marker using the diabetes marker high-sensitivity detection device according to any one of claims 1 to 8, comprising the steps of:
The high-pressure liquid chromatography pump provides a driving force to control the fluid, and a buffer solution carrying the sample is pumped into the droplet microfluidic chip from the sample injection valve so that the buffer solution drives the HbA1c sample to complete the separation of each component;
Wrapping the separated HbA1c sample components with an oil phase to create water-in-oil droplets;
Enabling the water-in-oil droplets to flow into a miniature detection pool, and detecting HbA1c sample components in the water-in-oil droplets in the miniature detection pool by using an ultraviolet spectrometer;
carrying out absorbance analysis on each component wrapped in the water-in-oil droplets by using an ultraviolet spectrometer to obtain a chromatogram of the separated component;
The HbA1c concentration ratio is obtained by calculating the peak area of HbA1c in the chromatogram to the peak area of total Hb by using data processing software of a computer.
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| CN201810572387.6A CN110568087A (en) | 2018-06-06 | 2018-06-06 | high-sensitivity detection device and method for diabetes marker |
| PCT/CN2019/090188 WO2019233446A1 (en) | 2018-06-06 | 2019-06-05 | Highly sensitive detection apparatus and method for diabetes marker |
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| CN102068409A (en) * | 2011-01-13 | 2011-05-25 | 清华大学 | Method for preparing mono-disperse microemulsion, liposome and microsphere based on microfluidic technology |
| NL2008483C2 (en) * | 2012-03-13 | 2013-09-16 | Univ Leiden | Method and device for solvent evaporation from a liquid feed. |
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| WO2016127073A1 (en) * | 2015-02-06 | 2016-08-11 | The Regents Of The University Of Michigan | Controlled coalescence of gas phase segmented droplets |
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2018
- 2018-06-06 CN CN201810572387.6A patent/CN110568087A/en active Pending
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2019
- 2019-06-05 WO PCT/CN2019/090188 patent/WO2019233446A1/en active Application Filing
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| WO2019233446A1 (en) | 2019-12-12 |
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Application publication date: 20191213 |