CN116448850A - Detection device for dopamine in tear, preparation method and application - Google Patents
Detection device for dopamine in tear, preparation method and application Download PDFInfo
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/301—Reference electrodes
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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Abstract
The present disclosure provides a detection device of dopamine in tear, comprising: the electrochemical chip (1), dopamine specific sensitive film (2) is loaded on the electrochemical chip to catalyze dopamine molecules in tear to generate oxidation reaction under the condition that voltage is applied to the electrochemical chip so as to enable the dopamine molecules to lose electrons, thereby generating response current in the electrochemical chip, and the shell (3) is provided with through holes, and the through holes are suitable for being combined with the electrochemical chip (1) to form an electrolytic cell suitable for containing tear. The disclosure also provides a construction method of the device for detecting dopamine in tear and a method for measuring the content of dopamine in tear.
Description
Technical Field
The present disclosure relates to electrochemical detection of dopamine, and more particularly to a device for detecting dopamine in tears, and a preparation method and application thereof.
Background
Dopamine (DA) is the most abundant neurotransmitter in the brain, and helps cells to transmit pulse chemicals to regulate nervous system activity in the human body, thus playing an important role in various processes such as learning, emotion, memory, motor control, endocrine system regulation and vision regulation. The abnormal level of dopamine is closely related to various diseases, such as eye diseases including myopia, glaucoma and the like, and systemic diseases including primary sicca syndrome, parkinsonism, schizophrenia, depression, alzheimer's disease and the like, and the dopamine is qualitatively and quantitatively analyzed, so that physiological changes can be monitored, disease mechanisms are insight, and a research foundation is laid for diagnosis and treatment of clinical diseases.
The tear sample is utilized to realize the detection of the in-vivo dopamine level, and the method has the characteristics of non-invasiveness, convenience in acquisition, safety in operation and the like. However, under normal conditions, the tear sample size is very small, with a single capillary pipette collection of only about 3 μl. Also, the concentration of dopamine in the tear sample may change rapidly over time.
Currently, common dopamine detection methods are high performance liquid chromatography and enzyme-linked immunosorbent assay. However, both of these detection techniques require a large volume of the detection sample, and require long pretreatment of the tear sample to exclude other interfering substances. Moreover, the related art detection system is complex, has long processing and analysis time, and cannot meet the requirements of portable, rapid and micro-tear sample detection.
Therefore, developing a device for detecting the content of dopamine in a trace sample of tears is of great importance for practical application and clinical analysis.
Disclosure of Invention
In view of the above, in order to solve at least one technical problem in the prior art and other aspects, the present disclosure provides a device for detecting dopamine in tear, a preparation method and an application thereof, establishes a quantitative relationship between the concentration of dopamine in tear and the intensity of response current, and realizes the detection of dopamine in different clinical tear samples.
In one aspect of the present disclosure, a device for detecting dopamine in tear fluid is disclosed, comprising:
an electrochemical chip; the dopamine specific sensitive film is loaded on the electrochemical chip to specifically catalyze dopamine molecules in tear to generate oxidation reaction under the condition that the electrochemical chip applies voltage so as to lead the dopamine molecules to lose electrons, thereby generating response current in the electrochemical chip; and a housing having a through-hole formed therein, the through-hole being adapted to be combined with the electrochemical chip to form an electrolytic cell adapted to contain a minute amount of tears.
According to an embodiment of the present disclosure, an electrochemical chip includes:
the working electrode is loaded with a dopamine specific sensitive film to capture and transmit electrons; the auxiliary electrode is suitable for forming a polarization loop with the working electrode; and a reference electrode adapted to provide a reference voltage to measure an electrode potential of the working electrode.
According to an embodiment of the present disclosure, the diameter of the through hole is less than 2mm.
According to an embodiment of the present disclosure, each of the working electrode, the auxiliary electrode, and the reference electrode is at least partially exposed in the through hole.
According to an embodiment of the present disclosure, the volume range of the electrolytic cell includes 2 to 5. Mu.L.
According to embodiments of the present disclosure, the voltage range includes 0.1V to 0.2V.
In another aspect of the present disclosure, a method for constructing a device for detecting dopamine in tear fluid described above is disclosed, including:
dissolving pyrrole, single-walled carbon nanotubes and gold nanoparticles in ultrapure water to obtain a mixed solution, and electrochemically depositing the mixed solution on a working electrode to obtain a dopamine specific sensitive film;
uniformly mixing an electrochemical corrosion resistant material and a curing agent, and after high-temperature curing, punching a through hole with the diameter smaller than 2mm in the interior to obtain a shell;
and (3) bonding the shell with an electrochemical chip after plasma treatment to obtain the device for detecting dopamine in tear.
According to the embodiment of the disclosure, the mass fraction range of pyrrole comprises 0.8% -4.0%; the mass fraction range of the single-wall carbon nano tube comprises 0.05% -0.2%; the mass fraction range of the gold nanoparticles is 0.0004% -0.002%.
According to embodiments of the present disclosure, the electrochemical deposition process conditions are deposition at a constant potential of 0.75V to 0.85V for 150s to 600s.
In another aspect of the present disclosure, a method for measuring the content of dopamine in tear fluid using the above-described device for detecting dopamine in tear fluid is disclosed.
According to the embodiment of the disclosure, a miniature electrochemical detection device is provided, a miniature electrolytic cell directly accommodating untreated micro tears is constructed by a shell and an electrochemical chip, and through electrochemical reaction, when dopamine molecules in tears are in contact with the electrochemical chip loaded with a dopamine specific sensitive film, the dopamine molecules lose electrons through applying a certain voltage so as to be oxidized into dopamine quinone. Meanwhile, the surface of the electrochemical chip captures the electrons and generates response current to be transmitted to an electrochemical workstation, so that the detection of the content of dopamine in tear is realized.
Drawings
FIG. 1 is a schematic diagram of a test of a device for detecting dopamine in tear fluid in the present disclosure;
FIG. 2 is an overall schematic diagram of a device for detecting dopamine in tear fluid in accordance with the present disclosure;
FIG. 3 is a schematic diagram of an electrochemical chip of the device for detecting dopamine in tear fluid of the present disclosure;
FIG. 4 is a schematic diagram of the electrochemical chip of the device for detecting dopamine in tear in the present disclosure in combination with a dopamine-specific sensitive film;
FIG. 5 is a flow chart of the preparation of a device for detecting dopamine in tear fluid in accordance with the present disclosure;
FIG. 6 is a graph of a fit of dopamine concentration versus current density for a device for detecting dopamine in tear fluid in accordance with the present disclosure;
FIG. 7 is a graph showing the effect of the device for detecting dopamine in tear fluid at various dopamine concentrations in the present disclosure;
FIG. 8 is a graph showing the comparison of the detection of dopamine in tears by the device for detecting dopamine in tears of the present disclosure;
FIG. 9 is a graph showing the comparison of the concentration of dopamine in tears before and after treatment by red eye irradiation, as detected by the dopamine in tears detection device in the test example of the present disclosure; and
fig. 10 is a graph of the dopamine in tear detection device monitoring changes in the concentration of dopamine in the daytime tear in the test examples of the present disclosure.
Reference numerals
An electrochemical chip 1;
a working electrode 1.1;
an auxiliary electrode 1.2;
a reference electrode 1.3;
a substrate 1.4;
a dopamine-specific sensitive film 2; and
a housing 3.
Detailed Description
For the purposes of promoting an understanding of the principles and advantages of the disclosure, reference will now be made to the embodiments illustrated in the drawings and specific language will be used to describe the same.
The endpoints of the ranges and any values disclosed in this disclosure are not limited to the precise range or value, and such range or value should be understood to encompass values approaching those range or value. For numerical ranges, one or more new numerical ranges may be obtained in combination with each other between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point value, and are to be considered as specifically disclosed in this disclosure.
The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the disclosure. The terms "comprises," "comprising," and/or the like, as used herein, specify the presence of stated features, steps, operations, and/or components, but do not preclude the presence or addition of one or more other features, steps, operations, or components.
All terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art unless otherwise defined. It should be noted that the terms used herein should be construed to have meanings consistent with the context of the present specification and should not be construed in an idealized or overly formal manner.
Compared with the high performance liquid chromatography and the enzyme-linked immunosorbent assay in the related technology, the electrochemical detection has the advantages of simple operation, rapid detection, portable instrument, lower cost and the like. However, the conventional electrochemical test device requires at least 5mL or more, which means that the collected micro-tear sample is diluted, which results in a change of the components in the tear fluid, resulting in poor accuracy of the test result.
Fig. 1 is a schematic diagram of a test of a device for detecting dopamine in tear fluid in the present disclosure.
As shown in fig. 1, in the present disclosure, a device for detecting dopamine in tear fluid applies a microfluidic technology to set an electrolytic cell structure according to the property of a tiny fluid (volume range is 2-5 μl) of tear fluid, and a dopamine specific sensitive film is loaded on an electrochemical chip, so that electrons are lost by catalytic oxidation of dopamine molecules in tear fluid through an electrochemical method, and then the electrons are received through an electrode to output corresponding current and transmitted to an electrochemical workstation externally connected with the detection device. In the subsequent treatment process, according to Faraday's law, a quantitative relation between the concentration of dopamine and the intensity of response current can be established, so that the quantitative detection of the dopamine is realized.
The present disclosure pertains to "Microfluidics" and refers to science and technology related to systems that use microchannels (tens to hundreds of microns in size) to process or manipulate minute fluids, and features such as miniaturization and integration.
Fig. 2 is an overall schematic diagram of a device for detecting dopamine in tear in the present disclosure.
In one aspect of the present disclosure, a device for detecting dopamine in tear fluid is disclosed, as shown in fig. 2, comprising an electrochemical chip 1, a dopamine-specific sensitive film 2, and a housing 3. The electrochemical chip 1 applies a voltage to cause oxidation of the dopamine molecules and to generate a response current. The dopamine specific sensitive film 2 is loaded on the electrochemical chip to catalyze dopamine molecules in tear to generate oxidation reaction under the condition that the electrochemical chip applies voltage so as to enable the dopamine molecules to lose electrons, thereby generating response current in the electrochemical chip. The housing 3 is formed with a through-hole adapted to form an electrolytic cell adapted to contain tears in combination with the electrochemical chip 1.
According to an embodiment of the present disclosure, a micro electrochemical detection device is provided in the present disclosure, in which a micro electrolytic cell directly accommodating untreated micro tears is constructed by a housing and an electrochemical chip, and through electrochemical reaction, when dopamine molecules in tears are in contact with the electrochemical chip loaded with a dopamine specific sensitive film, the dopamine molecules are catalyzed to lose electrons by applying a certain voltage, so that they are oxidized into dopamine quinone. At the same time, the electrochemical chip surface captures these electrons and generates a response current for transmission to the electrochemical workstation by measuring the response current intensity during the reaction. Then, according to Faraday's law, a quantitative relation between the concentration of dopamine and the intensity of response current can be established, and the detection of the content of dopamine in tear is realized. The miniature electrochemical detection device provided by the disclosure has the advantages of simple operation, portability and accurate quantitative detection for a trace sample.
Fig. 3 is a schematic diagram of an electrochemical chip of the device for detecting dopamine in tear in the present disclosure.
According to an embodiment of the present disclosure, as shown in fig. 3, the electrochemical chip 1 includes a substrate 1.4, and a working electrode 1.1, an auxiliary electrode 1.2, and a reference electrode 1.3 juxtaposed on the substrate 1.4.
Fig. 4 is a schematic diagram of the electrochemical chip and dopamine-specific sensitive thin film binding of the device for detecting dopamine in tear in the present disclosure.
As shown in fig. 4, the working electrode 1.1 is loaded with a dopamine-specific sensitive film 2 to capture and transport electrons; the auxiliary electrode 1.2 is suitable for forming a polarization loop with the working electrode 1.1; the reference electrode 1.3 is adapted to provide a reference voltage to measure the electrode potential of the working electrode 1.1.
According to the embodiment of the disclosure, on an electrochemical chip, a working electrode 1.1 is used for extracting and outputting an electric signal in a detection device, and the electric signal is transmitted to an external electrochemical workstation through an auxiliary electrode 1.2 for processing, wherein a reference electrode 1.3 is used as a standard for auxiliary measurement of electrode potential information of the working electrode.
According to an embodiment of the present disclosure, wherein the working electrode 1.1 may be selected to be an indium tin oxide material, the auxiliary electrode 1.2 may be selected to be a metallic platinum material, and the reference electrode 1.3 may be selected to be a silver/silver chloride material.
According to embodiments of the present disclosure, the diameter of the through hole is less than 2mm, e.g., 0.5mm, 1mm, 1.5mm, etc.
According to the embodiments of the present disclosure, since the volume of tear is small, in combination with the microfluidic technology, it is necessary to construct a micro-cavity.
According to an embodiment of the present disclosure, each of the working electrode 1.1, the auxiliary electrode 1.2 and the reference electrode 1.3 is at least partially exposed in the through hole.
According to embodiments of the present disclosure, the working electrode 1.1, the auxiliary electrode 1.2 and the reference electrode 1.3 in the electrochemical chip need to be simultaneously contacted with tear fluid for a cooperative assay.
According to embodiments of the present disclosure, the volume range of the electrolytic cell includes 2 to 5. Mu.L, for example, 2. Mu.L, 2.5. Mu.L, 3. Mu.L, 3.5. Mu.L, 4. Mu.L, 4.5. Mu.L, 5. Mu.L, etc.
According to the embodiment of the present disclosure, the volume of the electrolytic cell satisfies the requirement of containing a minute amount of tear in this range.
According to embodiments of the present disclosure, the applied voltage range includes 0.1V to 0.2V, for example, 0.1V, 0.13V, 0.15V, 0.18V, 0.2V, and the like.
According to the embodiment of the disclosure, the detection device provided by the disclosure needs to generate electrochemical reaction through externally applied potential or current, 0.1V-0.2V is the specific oxidation potential of the dopamine molecules, and when the externally applied voltage range is within the range, the dopamine molecules can be directionally catalyzed and oxidized to shield the influence of other interfering substances in the detection sample, so that the specific detection of the dopamine is realized.
Fig. 5 is a flow chart of the preparation of a device for detecting dopamine in tear fluid in accordance with the present disclosure.
In another aspect of the present disclosure, a method for constructing a device for detecting dopamine in tear fluid described above is disclosed, as shown in fig. 5, including:
s1: dissolving pyrrole, single-walled carbon nanotubes and gold nanoparticles in ultrapure water to obtain a mixed solution, and electrochemically depositing the mixed solution on a working electrode 1.1 to obtain a dopamine specific sensitive film 2;
s2: uniformly mixing an electrochemical corrosion resistant material and a curing agent, and after high-temperature curing, punching a through hole with the diameter smaller than 2mm in the interior to obtain a shell 3;
s3: and bonding the shell 3 with the electrochemical chip 1 after plasma treatment to obtain the device for detecting dopamine in tear.
According to the embodiment of the disclosure, the pyrrole and the single-walled carbon nanotube can form a layer of organic composite material film with large specific surface area and strong conductivity on the surface of the working electrode 1.1 so as to enhance the catalytic performance, and the introduction of the gold nanoparticles further enhances the electron transfer rate and the catalytic performance in the electrochemical process and improves the detection performance of the electrochemical detection device.
According to embodiments of the present disclosure, the mass fraction of pyrrole ranges from 0.8% to 4.0%, such as 0.8%, 1.0%, 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, etc.; the mass fraction of the single-walled carbon nanotubes ranges from 0.05% to 0.2%, for example, from 0.05%, 0.08%, 0.1%, 0.12%, 0.15%, 0.18%, 0.2%, etc.; the mass fraction of the gold nanoparticles ranges from 0.0004% to 0.002%, for example, from 0.0004%, 0.0006%, 0.0008%, 0.001%, 0.0012%, 0.0014%, 0.0016%, 0.0018%, 0.002%, and the like.
According to the embodiment of the present disclosure, through a plurality of experiments, the dopamine-specific sensitive film 2 with optimal performance can be obtained under the condition of limiting the components so as to catalyze the oxidation of dopamine.
According to embodiments of the present disclosure, the electrochemical deposition process conditions are deposition at a constant potential of 0.75V to 0.85V for 150s to 600s, such as deposition for 150s, 200s, 250s, 300s, 350s, 400s, 450s, 500s, 550s, 600s.
According to the embodiment of the disclosure, through a large number of experiments, the dopamine-specific sensitive film 2 with good appearance and thickness suitable for detection can be obtained under the limited condition of electrochemical deposition so as to catalyze the oxidation of dopamine
In another aspect of the present disclosure, a method for measuring the content of dopamine in tear fluid using the above-described device for detecting dopamine in tear fluid is disclosed.
Fig. 6 is a graph showing a relationship between the concentration of dopamine in a tear and the current density in the detection device of dopamine in tear in the present disclosure.
Fig. 7 is a graph showing the effect of the device for detecting dopamine in tear in the present disclosure at different dopamine concentrations.
As shown in fig. 6 and 7, the detection device for dopamine in tear provided in the present disclosure can detect 1nM to 250nM dopamine molecules by processing electric signals and faraday's law, and has a response time of about 3s, and performs a good linear regression fit, so that the detection device has the effects of high efficiency, sensitivity and accurate quantitative measurement.
According to the embodiment of the disclosure, the response current of the detection device for the dopamine in the tear still shows a good linear relation with the concentration of the dopamine in the presence of various interferents, so that the concentration of the trace dopamine in the tear can be accurately determined through the magnitude of the response current. The detection device of dopamine in tear in the present disclosure is of great significance in revealing the intrinsic relation between eye diseases and the concentration of dopamine in tear.
It should be noted that the described embodiments are only some embodiments of the present disclosure, and not all embodiments. Based on the embodiments in this disclosure, other embodiments that may be obtained by one of ordinary skill in the art without making any inventive effort are within the scope of the present disclosure.
Examples
S1: the electrochemical chip 1 was sequentially subjected to ultrasonic cleaning in acetone, absolute ethanol and ultrapure water for 5 minutes and dried in an air atmosphere. The mixed solution is prepared from ultrapure water, wherein the mixed solution comprises about 0.1% of pyrrole, about 0.05% of single-walled carbon nano-tubes and about 0.0004% of gold nano-particles by mass fraction. Indium tin oxide is used as a working electrode, a platinum wire is used as an auxiliary electrode, silver/silver chloride is used as a reference electrode, and the electrochemical chip 1 loaded with the dopamine-specific sensitive film 2 is obtained by depositing 150s-600s at a constant potential of about 0.85V and drying in air.
S2: polydimethylsiloxane (PDMS) was mixed with the curing agent solution in a ratio of 10:1, and placed in a freeze dryer to remove air bubbles by vacuum. Then, the mixture was put into an oven at about 90℃for baking and curing for about 90 minutes to obtain a PDMS film having a thickness of about 1mm, and a through hole having a diameter of 2mm was bored in the inside to obtain a housing 3.
S3: the housing 3 was subjected to plasma treatment for about 2 minutes and then press-bonded to the electrochemical chip 1, as shown in fig. 2, to obtain a device for detecting dopamine in tear.
S4: the electrochemical chip 1 in the detection device for the dopamine in the tear is connected with an electrochemical workstation to measure the content of the dopamine in the tear.
Test case
An interferent solution with a set concentration is prepared, and electrochemical detection is carried out under the applied voltage range of 0.1V-0.2V. Wherein the interferent solution comprises uric acid, glucose, ascorbic acid, albumin, sodium chloride and potassium chloride.
Fig. 8 is a graph showing the comparison of the detection of interferents and dopamine in tears by the device for detecting dopamine in tears of the present disclosure.
As shown in fig. 8, the response of the device to interferents such as uric acid, glucose, ascorbic acid, albumin, sodium chloride and potassium chloride is far smaller than that of dopamine, which proves that the microporous electrochemical device can perform specific detection of dopamine molecules in the presence of common tear interferents.
Application example 1
Red eye irradiation is a common myopia treatment means in hospitals at present. The detection device for the dopamine in the tears, which is prepared by the embodiment of the disclosure, is used for detecting the concentration change of the dopamine in the tears before and after the infrared eye irradiation treatment of a myopic patient.
The experimental group receives three-minute red-light eye irradiation treatment, the same batch of subjects is adopted for self control, the control group receives three-minute dark treatment, tear collection is performed immediately after the two groups of treatments, and the detection is performed through the detection device of dopamine in tear, which is prepared in the embodiment of the disclosure.
Fig. 9 is a graph showing the comparison of the concentration changes of dopamine in tears before and after treatment by red eye irradiation of a myopic patient detected by the dopamine detection device in tears in the test example of the present disclosure.
As shown in fig. 9, the red eye irradiation treatment was found to significantly increase the content of dopamine molecules in tears. The detection device for dopamine in tears can efficiently, sensitively and accurately detect the change of the concentration of the dopamine in tears, and verifies that the dopamine molecules can be stimulated to generate through red light irradiation so as to regulate the neural activity.
Application example 2
Knowing the concentration change of the dopamine content in human tears in one day has profound significance for knowing the neuromodulation activity of the human body.
The daytime curve of the concentration of the dopamine in the tears is monitored for experimental groups by using the detection device of the dopamine in the tears prepared in the embodiment of the disclosure. The test collects tears of the subjects at different moments in the day for a plurality of consecutive days for dopamine detection.
Fig. 10 is a graph of the dopamine in tear detection device monitoring changes in the concentration of dopamine in the daytime tear in the test examples of the present disclosure.
As shown in fig. 10, the change rule of the concentration of dopamine in tear can be obtained from the graph after detection, the concentration of dopamine in tear in human body in one day is in a trend of increasing and then decreasing, and the concentration of dopamine in tear in human body reaches a peak value at 2 pm. It is demonstrated that the device for detecting dopamine in tear fluid proposed in the present disclosure can be used to monitor the concentration of dopamine in tear fluid.
While the foregoing is directed to embodiments of the present disclosure, other and further details of the invention may be had by the present application, it is to be understood that the foregoing description is merely exemplary of the present disclosure and that no limitations are intended to the scope of the disclosure, except insofar as modifications, equivalents, improvements or modifications may be made without departing from the spirit and principles of the present disclosure.
Claims (10)
1. A device for detecting dopamine in tear fluid, comprising:
an electrochemical chip (1);
the dopamine specific sensitive film (2) is loaded on the electrochemical chip to catalyze dopamine molecules in tear to undergo oxidation reaction under the condition that voltage is applied to the electrochemical chip so as to enable the dopamine molecules to lose electrons, and thus response current is generated in the electrochemical chip; and
and a housing (3) on which a through hole is formed, said through hole being adapted to form, in combination with the electrochemical chip (1), an electrolytic cell adapted to contain the tear fluid.
2. The device according to claim 1, wherein the electrochemical chip (1) comprises, juxtaposed:
a working electrode (1.1) loaded with the dopamine-specific sensitive film (2) to capture and transport the electrons;
an auxiliary electrode (1.2) adapted to form a polarization circuit with the working electrode (1.1); and
a reference electrode (1.3) adapted to provide a reference voltage to measure the electrode potential of the working electrode (1.1).
3. The device of claim 1, wherein the diameter of the through hole is less than 2mm.
4. A device according to any one of claims 1 to 3, wherein each of the working electrode (1.1), the auxiliary electrode (1.2) and the reference electrode (1.3) is at least partially exposed in the through hole.
5. The apparatus of claim 1, wherein the volume of the electrolytic cell ranges from 2 to 5 μl.
6. The apparatus of claim 1, wherein the applied voltage range comprises 0.1V to 0.2V.
7. A method for constructing a device for detecting dopamine in tear fluid according to any one of claims 1 to 6, comprising:
dissolving pyrrole, single-walled carbon nanotubes and gold nanoparticles in ultrapure water to obtain a mixed solution, and electrochemically depositing the mixed solution on a working electrode (1.1) to obtain the dopamine specific sensitive film (2);
uniformly mixing an electrochemical corrosion resistant material with a curing agent, and after high-temperature curing, punching a through hole with the diameter smaller than 2mm in the interior to obtain the shell (3);
and bonding the shell (3) with the electrochemical chip (1) after plasma treatment to obtain the detection device of dopamine in tear.
8. The method of claim 7, wherein,
the mass fraction range of the pyrrole comprises 0.8% -4.0%;
the mass fraction range of the single-walled carbon nanotube comprises 0.05% -0.2%;
the mass fraction range of the gold nanoparticles comprises 0.0004% -0.002%.
9. The method of claim 7, wherein the electrochemical deposition process conditions are deposition at a constant potential of 0.75V to 0.85V for 150s to 600s.
10. A method of measuring the dopamine content of tears using a device according to any of claims 1 to 6.
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