WO2019233446A1

WO2019233446A1 - Highly sensitive detection apparatus and method for diabetes marker

Info

Publication number: WO2019233446A1
Application number: PCT/CN2019/090188
Authority: WO
Inventors: 赫斯奥夫·哈瑞; 穆桑特·卢卡; 马丁阿尔贝托·贝尼托; 萨拉斯瓦特·马扬克; 塔塔奇多洛塔·艾娃; 赫斯奥夫瑞塔·凯撒; 张贯京; 邹和群; 葛新科; 肖应芬; 唐小浪; 刘义
Original assignee: 深圳市贝沃德克生物技术研究院有限公司
Priority date: 2018-06-06
Filing date: 2019-06-05
Publication date: 2019-12-12
Also published as: CN110568087A

Abstract

A highly sensitive detection apparatus and method for a diabetes marker. The apparatus comprises a droplet-based micro-fluidic chip (1), a high pressure liquid chromatography pump (2), an injection valve (3), a deuterium tungsten halogen light source (10), an ultraviolet spectrometer (11), and a computer (13). The high pressure liquid chromatography pump (2) provides a driving pressure to pump a buffer solution into a pipe of the droplet-based micro-fluidic chip (1). The injection valve (3) introduces an HbA1c sample from a sample inlet (4) to complete quantitative injection. A strong cation exchange polymer monolithic column (7) of the droplet-based micro-fluidic chip (1) separates components of the HbA1c sample. A droplet generator (8) wraps the separated HbA1c components to form water-in-oil droplets and said droplets flow into a miniaturized detection cell (9). The deuterium tungsten halogen light source (10) provides ultraviolet light. The ultraviolet spectrometer (11) receives the ultraviolet light, measures the absorbance of the HbA1c sample, and transmits a detection signal to data processing software of the computer (13) by means of an optical fiber (12) for data processing to obtain an HbA1c concentration ratio. Such an apparatus and method not only improve the HbA1c detection sensitivity, but also are easy to operate and beneficial to field real-time detection.

Description

Device and method for detecting diabetes markers with high sensitivity

Technical field

The invention relates to the technical field of detection of diabetes markers, in particular to a device and method for detecting diabetes markers with high sensitivity.

Background technique

Diabetes is a group of metabolic diseases characterized by high blood sugar, with a high incidence. It is one of the four major diseases that currently endanger human health. Long-term diabetes is likely to cause complications such as diabetic ketones, cardiovascular disease, neuropathy, diabetic foot, and diabetic nephropathy, which seriously threatens human health. According to the Diabetes Branch of the Chinese Medical Association in 2013, it is estimated that there are 139 million diabetic patients in China, of which 60.7% are not diagnosed.

For many years, the diagnosis of diabetes has been based on plasma glucose levels, including fasting blood glucose and oral glucose tolerance tests for 2 hours. However, blood glucose testing methods are susceptible to factors such as diet, exercise, and medication, as well as the need to take blood for a specific time, which greatly limits the accuracy of the test results and brings inconvenience to patients. There are two main types of HbA1c detection methods currently used in clinical laboratories: one is based on the different charges of glycated hemoglobin and non-glycated hemoglobin, such as ion exchange chromatography, electrophoresis, and other methods; another method These methods are based on the structural characteristics of glycated groups on hemoglobin, such as affinity chromatography, ion capture, and immunoassay. In 2002, the International Federation of Clinical Chemistry (IFCC) published the international standard method for HbA1c detection, that is, the 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, which makes it difficult to clinically promote it. At present, the ion exchange HPLC method is considered as the gold standard for the analysis and detection of HbA1c, which measures the percentage content of HbA1c in the total hemoglobin (Hb) in the blood. However, this method uses a commercial cation-exchange chromatography column combined with extra-column detection, which results in large reagent consumption and long analysis time, and a large dead volume between the end of the column and the detector due to the tubing connection. Separated sample components are susceptible to diffusion and remixing, reducing detection sensitivity. Therefore, the prior art has disadvantages such as low detection sensitivity, large detection instrument volume, and complicated operation steps.

technical problem

The main object of the present invention is to provide a device and method for detecting diabetes markers with high sensitivity, which aims at the technical problem of low sensitivity of detecting diabetes markers in the prior art.

Technical solutions

To achieve the above object, the present invention provides a high-sensitivity detection device for diabetes markers, including a droplet microfluidic chip, a high-pressure liquid chromatography pump, a sampling valve, a deuterium halogen lamp light source, an ultraviolet spectrometer, and a computer, wherein:

The high pressure liquid chromatography pump is used to provide driving pressure to pump the buffer solution into the pipeline of the droplet microfluidic chip; the sampling valve passes the HbA1c sample from the sample inlet and flows out from the sample waste liquid outlet Quantitative injection is completed; the droplet microfluidic chip includes a strong cation exchange polymer monolithic column, a droplet generator, and a micro detection cell; the strong cation exchange polymer monolithic column is used to separate groups of HbA1c samples The droplet generator is used for wrapping the separated HbA1c components to form a series of water-in-oil droplets with different concentrations; the miniature detection cell is aligned with the optical path of the ultraviolet spectrometer; the deuterium halogen lamp light source is used for Provide ultraviolet light for detecting absorbance of HbA1c samples; the ultraviolet spectrometer is used to receive ultraviolet light for detecting absorbance of HbA1c samples and transmit the detection signal to a computer through an optical fiber; the computer is provided with data processing software for The detection signal is subjected to data processing to obtain a HbA1c concentration ratio.

Preferably, the droplet microfluidic chip is composed of a cover sheet layer, an upper fluid channel layer, a lower fluid channel layer, and a base layer in this order.

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. The water phase inlet penetrates the cover sheet layer and is introduced into the water phase on the upper fluid channel layer. The passage is connected.

Preferably, the shape of the water phase introduction channel is a curved serpentine shape, and the inside of the water phase introduction channel is filled with a strong cation exchange polymer monolith.

Preferably, a chromatographic column is provided inside the water phase introduction channel, and the first oil phase inlet and the second oil phase inlet penetrating the cover sheet layer pass through the first oil phase introduction channel and the second oil phase on the upper fluid channel layer, respectively. A liquid droplet generator which is introduced into the channel and forms a cross-shaped channel with the water phase introduction channel and the first liquid droplet channel.

Preferably, the cross-shaped channel has a contraction structure with an arc shape, and the width of the cross-shaped channel satisfies the relationship 2 * W1≥W2≥0.5 * W1, 2 * W1≥W3 = W4≥0.5 * W1, and W1> W2 Where * indicates a multiplication sign, W1 is the width of the end channel of the water phase introduction channel, W2 is the width of the inlet channel of the first droplet channel, W3 is the width of the end channel of the first oil phase introduction channel, and W4 is the second oil phase introduction The end channel width of the channel, 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, a first small hole penetrating through the upper fluid channel layer and a second small hole penetrating through the lower fluid channel layer The positions are corresponding, and the micro detection cell is formed after the two are penetrated.

Preferably, the micro detection cell is in communication with the first liquid droplet channel on the upper fluid channel layer, and is in communication with the second liquid droplet channel penetrating through the lower fluid channel layer, and the second liquid droplet channel passes through the lower fluid channel in sequence. The fluid outlets of the upper layer, the upper fluid channel layer and the cover sheet layer realize circulation with the outside world.

Preferably, the droplet microfluidic chip is made of one or more of polymethyl methacrylate (PMMA), polycarbonate (PC), polystyrene (PS), and cycloolefin copolymer (COC). Made of several combined materials.

In another aspect, the present invention also provides a method for detecting a diabetes marker using the diabetes marker high-sensitivity detection device. The method includes the steps of: controlling a fluid by providing a driving force through a high-pressure liquid chromatography pump, and The sample buffer is pumped into the droplet microfluidic chip from the injection valve to make the buffer drive the HbA1c sample to complete the separation of the components; the separated HbA1c sample components are wrapped with the oil phase to generate water-in-oil droplets; The water-in-oil droplets flow into the micro-detection cell, and the HbA1c sample components in the water-in-oil droplets are detected by a UV spectrometer in the micro-detection cell; the ultraviolet-ray spectrometer is used to absorb the components wrapped in the water-in-oil droplets Analyze to obtain the chromatogram of the separated components; calculate the peak area of HbA1c in the chromatogram by the data processing software of the computer to the total area of Hb to obtain the HbA1c concentration ratio.

Beneficial effect

Compared with the prior art, the device and method for high sensitivity detection of diabetes markers according to the present invention adopts the above technical scheme, and achieves the following technical effects: The droplet microfluidic control for the device for high sensitivity detection of diabetes markers provided by the present invention The chip is small in size, low in cost, and simple in operation, which is conducive to real-time detection in the field. The invention integrates a cation exchange chromatographic column and a detection cell on a microfluidic chip, and adds a droplet generator between the end of the chromatographic column and the detection cell, which not only avoids the dead volume generated by the pipeline connection, but also uses the droplet's The detection method prevents the sample from diffusing and remixing at the end of the column, thus improving the detection sensitivity. In addition, the structure of the miniature detection cell is designed as a Z-type, which increases the optical path, thereby further improving the detection sensitivity. The droplet microfluidic chip is not only suitable for high-sensitivity detection of HbA1c, but also generally applicable to detection of biological macromolecules such as peptides, nucleic acids, and proteins with absorption peaks in the ultraviolet light band.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic structural diagram of a diabetes marker high-sensitivity detection device according to the present invention;

2 is a schematic structural diagram of a liquid droplet microfluidic chip of a device for detecting a diabetes marker of the present invention with high sensitivity;

3 is a schematic cross-sectional structure diagram of a droplet microfluidic chip of the diabetes marker high-sensitivity detection device of the present invention;

4 is a schematic structural diagram of a liquid droplet generator of a liquid droplet microfluidic chip of a high-sensitivity detection device for a diabetes marker according to the present invention;

FIG. 5 is a flowchart of a preferred embodiment of a detection method for a diabetes high-sensitivity detection device according to the present invention.

The realization of the purpose, functional characteristics and advantages of the present invention will be further described with reference to the embodiments and the drawings.

Embodiments of the invention

In order to further explain the technical means and effects adopted by the present invention to achieve the intended purpose of the present invention, the specific implementation, structure, features, and effects of the present invention are described in detail below with reference to the drawings and preferred embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present 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 high-sensitivity detection device for diabetes markers includes a droplet microfluidic chip 1, a high-pressure liquid chromatography pump 2, an injection valve 3, a deuterium 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 injection valve 3 passes the sample of glycated hemoglobin (HbA1c) from the sample inlet 4 And flow out from the sample waste liquid outlet 5 to complete the quantitative injection; the droplet microfluidic chip 1 includes a strong cation exchange polymer monolithic column 7, a droplet generator 8 and a micro detection cell 9; the strong cation exchange The polymer monolithic column 7 is used to separate the components in the HbA1c sample; the droplet generator 8 is used to wrap the separated HbA1c components to form a series of water-in-oil droplets with different concentrations; the micro-detection cell 9 Aligned with the optical path of the ultraviolet spectrometer 11; the deuterium-halogen tungsten light source 10 provides ultraviolet light for detecting the absorbance of the HbA1c sample; the ultraviolet spectrometer 11 is used for receiving the ultraviolet light for detecting the absorbance of the HbA1c sample and through the optical fiber 12 Detection signal transmission Input to computer 13; the computer 13 is installed with data processing software for data processing the detection signal to obtain the HbA1c concentration ratio.

Referring to FIG. 2, FIG. 2 is a schematic structural diagram of a droplet microfluidic chip of a high-sensitivity detection device for a diabetes marker according to 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 base layer 50 in this order; an aqueous phase is provided on the cover sheet layer 20 The inlet 21, the first oil phase inlet 22, the second oil phase inlet 23, and the fluid outlet 24. The water phase inlet 21 penetrates the cover sheet layer 20 and communicates with the water phase introduction channel 31 on the upper fluid channel layer 30; a chromatographic column is arranged inside the water phase introduction channel 31; and the first oil phase penetrating the cover film layer 20 The inlet 22 and the second oil phase inlet 23 pass through the first oil phase introduction channel 32 and the second oil phase introduction channel 33 on the upper fluid channel layer 30, respectively, and the water phase introduction channel 31 and the first liquid droplet channel 34 A droplet generator 8 forming a cross-shaped channel. The droplet generator 8 is formed by a first oil phase introduction channel 32 and a second oil phase introduction channel 33, and a water phase introduction channel 31 and a first liquid droplet channel 34. Cross-shaped passage.

As a preferred manner, the shape of the water phase introduction channel 31 is a curved serpentine shape, and the inside thereof is filled with a strong cation exchange polymer monolithic column. The strong cation exchange polymer monolithic column is a chromatographic column and its material is acrylic acid. Ester copolymer, loose and porous inside; the strong cation exchange polymer monolithic column is formed by light or thermally initiated polymerization. The mechanical strength, pore size, pH tolerance range, exchange capacity and other performance parameters of the monolithic column can be changed by changing the polymer. To adjust the type and ratio. The overall column channel length can be set in the range of 1 to 25 cm, the channel depth can be set in the range of 0.05 mm to 1 mm, and the channel width can be set in the range of 0.05 mm to 2 mm. The invention provides a chromatographic separation column with a loose and porous surface, low back pressure, large specific surface area, high column capacity and good permeability. Moreover, the column parameters of the chromatographic column such as the pore size and the particle size of the stationary phase can be controlled by prepolymerization. The type, proportion and triggering conditions of the objects are controlled to achieve the separation effect of different functions. The main functional components of the provided strong cation exchange polymer monolithic chip chip are composed of a polymer monolithic column, a droplet generator 8 and a micro detection cell 9. The size and pore structure of the polymer monolithic column can be changed according to different needs in practical applications. The width of the droplet generator 8 and the effective length of the micro-detection cell 9.

Referring to FIG. 3, FIG. 3 is a schematic cross-sectional structure diagram of a liquid droplet microfluidic chip of a high-sensitivity detection device for a diabetes marker according to the present invention. In this embodiment, the positions of 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 correspond to each other. After the two are penetrated, a miniature detection cell 9 is formed. The structural design is Z type. The micro-detection cell 9 communicates with the first liquid droplet channel 34 on the upper fluid channel layer 30 and communicates with the second liquid droplet channel 42 passing through the lower fluid channel layer 40; the second liquid droplet channel 42 passes through the lower fluid in sequence. The channel layer 40, the upper fluid channel layer 30, and the fluid outlet 24 of the cover sheet layer 20 communicate with the outside world.

Referring to FIG. 4, FIG. 4 is a schematic structural diagram of a droplet generator of a droplet microfluidic chip of a high-sensitivity detection device for a diabetes marker according to the present invention. As a preferred mode, the droplet generator 8 is a cross-shaped channel formed by the first oil phase introduction channel 32 and the second oil phase introduction channel 33, and the water phase introduction channel 31 and the first liquid droplet channel 34. The cross-shaped channel has an arc-shaped contraction structure, which is beneficial to the generation of small-sized droplets, and its width satisfies the relationship 2 * W1≥W2≥0.5 * W1, 2 * W1≥W3 = W4≥0.5 * W1, and W1> W2; where * represents the multiplication sign, W1 is the width of the end channel of the water phase introduction channel 31, W2 is the width of the inlet channel of the first droplet channel 34, W3 is the width of the end channel of the first oil phase introduction channel 32, W4 Is the width of the end channel of the second oil phase introduction channel 33, W is the width of the first droplet channel 34; its working principle is: use an external pump or valve drive system to inject the oil phase and water phase solutions from their respective inlets. At the intersection of the cross channel, the water-in-oil (W / O) droplets are generated by the interaction of the shear, viscous, surface tension, and inertial forces due to the compression of the water phase by the oil phase, and are The oil phase is carried into the first liquid droplet passage 34. The size of the liquid droplet generated by the liquid droplet generator 8 can be changed by adjusting the flow velocity ratio of the fluid in the first oil phase introduction channel 32 and the second oil phase introduction channel 33 and the water phase introduction channel 31.

In this embodiment, the droplet microfluidic chip 1 may be made of a transparent thermoplastic material, such as: polymethyl methacrylate (PMMA), polycarbonate (PC), polystyrene (PS), Cyclic olefin copolymer (COC) is made of one or several combination materials. The present invention illustrates the structure and dimensions of the liquid droplet microfluidic chip 1 made of different materials in the following specific embodiments.

As a first preferred embodiment, the material of the droplet microfluidic chip 1 is PMMA. The droplet microfluidic chip 1 includes four simple planar structures: a cover sheet layer 20, an upper fluid channel layer 30, and a lower fluid. The channel layer 40 and the base layer 50. The cover sheet layer 20 and the base layer 50 are both 2 mm thick, the upper fluid channel layer 30 and the lower fluid channel layer 40 are each 1 mm thick, and the overall polymer column is about 50 mm long, 0.5 mm wide, and 0.5 mm high; droplets occur. The cross channel of the device 8 is 0.5mm high, W1 = W2 = W3 = W4 = 0.2mm, W = 0.5mm; Miniature detection cell 9 diameter 1mm, height 2mm; water phase inlet 21, first oil phase inlet 22, second oil phase The inlet 23 and the fluid outlet 24 are each 1.6 mm in diameter.

As a second preferred embodiment, the material of the droplet microfluidic chip 1 is COC. The thickness of the cover sheet layer 20 and the base layer 50 of the droplet microfluidic chip 1 are both 2 mm, and the upper fluid channel layer 30 and the bottom The thickness of the fluid channel layer 40 is 0.5mm, the overall polymer column is about 30mm long, 0.5mm wide, and 0.3mm high; the droplet generator 8 cross channel is 0.3mm high, W1 = W2 = W3 = W4 = 0.3mm, W = 0.5 mm; the diameter of the miniature detection cell 9 is 0.6 mm and the height is 1 mm; the diameter of the water phase inlet 21, the first oil phase inlet 22, the second oil phase inlet 23 and the fluid outlet 24 are 1.0 mm.

In the droplet microfluidic chip 1 of the present invention, a buffer carrying a sample is introduced from the aqueous phase inlet 11 into a curved serpentine-shaped water phase introduction channel 31, and the curved serpentine structure can increase Under the condition of the length of the flow control chip, increasing the length of the overall polymer column in the aqueous phase introduction channel 31 is beneficial to the high-throughput separation and analysis of the sample. Moreover, the polymer monolithic column has the advantages of wide pH range tolerance, small pressure drop, high porosity, high exchange capacity, etc., which greatly reduces the requirements for high pressure output from the driving device, and has more choices for the mobile phase. In addition, polymer monolithic columns with different pore sizes and specific surface areas can be prepared by adjusting the ratio of the prepolymers, which is beneficial to improving the separation efficiency and speed. After the sample has been subjected to chromatographic separation in the polymer monolithic column, the oil solution injected from the first oil phase introduction channel 32 and the second oil phase introduction channel 33 is under the action of shear force, surface tension, viscous force and inertial force, respectively. Wrap the sample components flowing out of the end of the column in the form of droplets. This helps to ensure that the separated components do not undergo molecular diffusion before entering the micro-detection cell 9, and avoids the band broadening effect and the separated components to re- Mixing improves separation. In addition, integrating the micro-detection cell 9 on the droplet microfluidic chip facilitates real-time, online monitoring of the absorbance of the separated components, so as to obtain the real-time concentration of the sample, and the micro-detection cell 9 penetrates the upper fluid channel layer 30 and the bottom The fluid channel layer 40 increases the optical path length of the micro detection cell 9 and realizes a fully transparent detection cell, which greatly improves the detection sensitivity.

On the other hand, the present invention also provides a detection method based on the above-mentioned diabetes high-sensitivity detection device. Referring to FIG. 5, FIG. 5 is a flowchart of a preferred embodiment of a detection method for a diabetes high-sensitivity detection device according to the present invention. The method includes the following steps:

In step S51, the fluid is controlled by the driving force provided by the high-pressure liquid chromatography pump 2 and the buffer solution carrying the sample is pumped from the injection valve 3 into the droplet microfluidic chip 1 so that the buffer solution drives the HbA1c sample to complete each component. Separation. In this embodiment, the fluid is controlled by an external pump or a valve to provide a driving force, and the buffer carrying the sample is pumped from the water phase inlet 21 into the water phase introduction channel 31. Because different components in the sample and the water phase are introduced into the channel The force of the polymer monolithic column in 31 is different, and the moving speed of the sample in the polymer monolithic column is different. The separated components sequentially flow out of the polymer monolithic column.

In step S52, the separated HbA1c sample components are wrapped with an oil phase to generate water-in-oil droplets. In the present embodiment, the oil solution is separated from the ends of the water phase introduction channel 31 through the first oil phase introduction channel 32 and the second oil phase introduction channel 33 through the first oil phase inlet 22 and the second oil phase inlet 23, respectively. The resulting component aqueous solutions meet at the cross-shaped channel, and by controlling the flow rate ratio of the oil solution and the aqueous solution, water-in-oil droplets with a controllable particle size are formed and enter the first droplet channel 34.

Step S53, the water-in-oil droplets are flowed into the micro-detection cell 9, and the HbA1c sample component in the water-in-oil droplets is detected by the ultraviolet spectrometer 11 in the micro-detection cell 9; finally, the water-in-oil droplets subjected to absorbance detection From the fluid outlet 24 via the second droplet channel 42.

In step S54, absorbance analysis is performed on each component encapsulated in the water-in-oil droplets to obtain an ion chromatogram based on the droplet detection, that is, a chromatogram of the separated components. The ultraviolet spectrometer 11 is used to receive ultraviolet light and transmit signals to the data processing software of the computer 13 through the optical fiber 12 for data processing. Finally, the absorbance detection is completed, and the components wrapped in the water-in-oil droplets are absorbent. Analyze to get an ion chromatogram based on droplet detection.

In step S55, the HbA1c concentration ratio is obtained by calculating the peak area of HbA1c in the chromatogram and the peak area of the total Hb. In this embodiment, the data processing software of the computer 13 obtains the HbA1c concentration ratio value by calculating the peak area of the HbA1c in the chromatogram and the peak area of the total Hb (hemoglobin).

The liquid droplet microfluidic chip in the high-sensitivity detection device for diabetes markers provided by the present invention is small in volume, low in cost, simple in operation, and convenient for real-time detection in the field. In addition, the present invention also proposes a high-sensitivity detection method for diabetes markers. The method is not only suitable for high-sensitivity detection of HbA1c samples, but also for detection of biological macromolecules such as peptides, nucleic acids, and proteins. In order to improve the detection sensitivity, the present invention integrates a cation exchange chromatography column and a detection cell on a microfluidic chip, and adds a droplet generator between the end of the chromatographic column and the detection cell. This method not only avoids the death caused by the pipeline connection. The volume and detection in the form of droplets avoid the diffusion and remixing of the sample at the end of the column, thus improving the detection sensitivity. In addition, the structure of the miniature detection cell is designed as a Z-type, which increases the optical path, thereby further improving the detection sensitivity. The droplet microfluidic chip is not only suitable for high-sensitivity detection of HbA1c, but also generally applicable to detection of biological macromolecules such as peptides, nucleic acids, and proteins with absorption peaks in the ultraviolet light band.

The above are only preferred embodiments of the present invention, and thus do not limit the patent scope of the present invention. Any equivalent structure or equivalent process transformation made by using the description and drawings of the present invention, or directly or indirectly used in other related technical fields All are included in the patent protection scope of the present invention.

Industrial applicability

Claims

A high-sensitivity detection device for a diabetes marker is characterized in that the high-sensitivity detection device for a diabetes marker includes a droplet microfluidic chip, a high-pressure liquid chromatography pump, a sampling valve, a deuterium halogen lamp light source, an ultraviolet spectrometer, and Computer, where:

The high-pressure liquid chromatography pump is used to provide a driving pressure for pumping a buffer solution into a pipeline of a droplet microfluidic chip;

The sampling valve passes the HbA1c sample from the sample inlet and flows out from the sample waste liquid outlet to complete the quantitative sampling;

The droplet microfluidic chip includes a strong cation exchange polymer monolithic column, a droplet generator, and a micro detection cell; the strong cation exchange polymer monolithic column is used to separate components in a HbA1c sample; the droplet The generator is used for wrapping the separated HbA1c components to form a series of water-in-oil droplets with different concentrations; the miniature detection cell is aligned with the optical path of the ultraviolet spectrometer;

The deuterium halide tungsten light source is used to provide ultraviolet light for detecting absorbance of HbA1c samples;

The ultraviolet spectrometer is used for receiving ultraviolet light to perform absorbance detection on HbA1c samples and transmitting a detection signal to a computer through an optical fiber;

The computer is provided with data processing software for data processing the detection signal to obtain a HbA1c concentration ratio.
The high-sensitivity detection device for a diabetes marker according to claim 1, wherein the droplet microfluidic chip is composed of a cover sheet layer, an upper fluid channel layer, a lower fluid channel layer, and a base layer in this order.
The high-sensitivity detection device for a diabetes marker 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 The inlet penetrates the cover sheet layer and communicates with the water phase introduction channel on the upper fluid channel layer.
The high-sensitivity detection device for a diabetes marker according to claim 3, wherein the shape of the water phase introduction channel is a curved snake shape, and the inside of the water phase introduction channel is filled with a strong cation exchange polymer monolithic column.
The high-sensitivity detection device for a diabetes marker according to claim 3, wherein a chromatographic column is arranged inside the water phase introduction channel, and the first oil phase inlet and the second oil phase inlet passing through the cover sheet layer pass through the upper phase, respectively. The first oil phase introduction channel and the second oil phase introduction channel on the fluid channel layer form a droplet generator that forms a cross-shaped channel with the water phase introduction channel and the first liquid droplet channel.
The high-sensitivity detection device for diabetes markers according to claim 3, wherein the cross-shaped channel is a contraction structure with an arc shape, and the width of the cross-shaped channel satisfies the relationship 2 * W1≥W2≥0.5 * W1 , 2 * W1≥W3 = W4≥0.5 * W1 and W1> W2, where * represents a multiplication sign, W1 is the width of the end channel of the water phase introduction channel, W2 is the width of the inlet channel of the first droplet channel, and W3 is the first The end channel width of the oil phase introduction channel, W4 is the end channel width of the second oil phase introduction channel, and W is the front channel width of the first droplet channel.
The high sensitivity detection device for a diabetes marker according to claim 3, wherein the upper fluid channel layer is provided with a first small hole, and the lower fluid channel layer is provided with a second small hole, penetrating the upper fluid channel layer The positions of the first small hole and the second small hole penetrating the lower fluid channel layer correspond to each other, and the micro detection cell is formed after the two holes are penetrated.
The high-sensitivity detection device for a diabetes marker according to claim 7, wherein the micro-detection cell is in communication with the first liquid droplet channel on the upper fluid channel layer and with the second liquid droplet passing through the lower fluid channel layer The channels communicate with each other, and the second liquid droplet channel communicates with the outside world through a fluid outlet that sequentially penetrates the lower fluid channel layer, the upper fluid channel layer, and the cover sheet layer.
The high-sensitivity detection device for diabetes markers according to any one of claims 1 to 8, wherein the droplet microfluidic chip is made of polymethyl methacrylate, polycarbonate, polystyrene, ring The olefin copolymer is made of one or several combination materials.
A method for detecting a diabetes marker using the diabetes marker high-sensitivity detection device according to any one of claims 1 to 8, wherein the method includes the steps:

The high-pressure liquid chromatography pump provides driving force to control the fluid, and the buffer solution carrying the sample is pumped into the droplet microfluidic chip from the sampling valve to make the buffer drive the HbA1c sample to complete the separation of the components;

Encapsulating the separated HbA1c sample components with an oil phase to generate water-in-oil droplets;

The water-in-oil droplets are flowed into the micro-detection cell, and the HbA1c sample component in the water-in-oil droplets is detected by a UV spectrometer in the micro-detection cell;

Use ultraviolet light spectrometer to perform absorbance analysis on each component wrapped in water-in-oil droplets to obtain the chromatogram of the separated components;

The peak area of HbA1c in the chromatogram was calculated by the data processing software of the computer to obtain the HbA1c concentration ratio.