CN116519754A - Biochemical molecule detection method based on quantum capacitance effect of conductive composite material - Google Patents
Biochemical molecule detection method based on quantum capacitance effect of conductive composite material 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
- G01N27/02—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
- G01N27/22—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance
-
- 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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A20/00—Water conservation; Efficient water supply; Efficient water use
- Y02A20/20—Controlling water pollution; Waste water treatment
Abstract
The invention discloses a biochemical molecule detection method based on a quantum capacitance effect of a conductive composite material, belonging to the technical field of molecular sensors. Testing by adopting a three-electrode system of a working electrode, a reference electrode and a counter electrode, and enabling the working electrode and the reference electrode to be in contact with the solution to form a loop by dripping the solution into a liquid tank; and carrying out quantum capacitance test by using a lock-in amplifier or an impedance analyzer, evaluating the liquid environment of the sensing device according to the test result, and detecting the concentration of biochemical molecules in the liquid. The invention has the advantages of convenient preparation, controllable structure, high stability and good biocompatibility, can realize the instant sensing and information communication of the Internet, and has extremely high detection sensitivity.
Description
Technical Field
The invention relates to the technical field of molecular sensors, in particular to a biochemical molecular detection method based on a quantum capacitance effect of a conductive composite material.
Background
Biochemical molecular sensing technology plays an important role in the fields of biological medicine, environmental monitoring, clinical treatment, chemical industry and the like. The two-dimensional conductive material has excellent sensing sensitivity due to excellent electrical property, unique two-dimensional structure and good chemical inertness, and has great potential in the technical field of molecular sensing.
Quantum capacitance detection is used as a high-sensitivity detection mechanism, has unique advantages in detection application of target molecules in medical treatment and environment, and the sensing principle is based on detection of charges of target biochemical molecules. The quantum capacitance change signal reflects the significant change in energy state density caused by the field effect when charged biochemical molecules are adsorbed on the graphene surface. The biochemical molecules are adsorbed on the surface of the graphene quickly, and meanwhile, the electric signal detection has real-time property, so that the detection time is short, and the online quick detection can be realized. In the solid material, the two-dimensional material such as graphene is unique in that all atoms are positioned on the surface, so that the surface and the energy state density of the surface are extremely sensitive to environmental changes, the detection sensitivity is high, and trace detection can be realized. In 2009, graphene quantum capacitance detection is realized through ionic liquid, and so far, gas detection, pH value in solution, glucose detection and the like based on the graphene quantum capacitance have been realized. However, due to the current preparation and transfer technology of large-area two-dimensional materials still being immature, it is still difficult to obtain a highly stable and uniform two-dimensional substrate. In fact, current biochemical sensors based on conductive two-dimensional materials are mostly limited to laboratory testing. By compounding the conductive two-dimensional material with other materials, it would be one way to solve the above problems to obtain an inexpensive, stable multilayer composite material that can be free-standing in the macroscopic dimension (micrometer-scale thickness).
In general, sensing occurs only in a few layers of the surface nanolayers of the material, whereas the composite itself has a thickness on the order of microns, making the resistance change caused by biochemical molecular adsorption more difficult to detect. In fact, the maximum value of the thickness of the materials used for the gas and ion sensors reported today is only 15nm. Therefore, it is necessary to explore a biochemical molecular detection method of a conductive composite material with a macroscopic dimension of a micrometer-scale thickness. The conductive composite material is expected to be used for biochemical sensing of the surfaces of ceramic teeth, bones or other ceramic or polymer material products, and promotes the intelligent application of the products.
Disclosure of Invention
The invention aims at providing a biochemical molecule detection method based on a quantum capacitance effect of a conductive composite material, which is characterized by comprising the following steps of:
testing by adopting a three-electrode system of a working electrode, a reference electrode and a counter electrode, and enabling the working electrode and the reference electrode to be in contact with the solution to form a loop by dripping the solution into a liquid tank;
and carrying out quantum capacitance test by using a lock-in amplifier or an impedance analyzer, evaluating the liquid environment of the sensing device according to the test result, and detecting the concentration of biochemical molecules in the liquid.
The working electrode is made of conductive composite material, the reference electrode is calomel or silver chloride, and the counter electrode is graphite or platinum electrode.
The biochemical molecules comprise hydrogen ions, hydroxide ions, potassium ions, chloride ions, DNA, RNA and glucose.
The quantum capacitance test by using the phase-locked amplifier or the impedance analyzer is specifically as follows:
under the condition that the external direct current potential is unchanged, continuously changing the frequency of the external alternating current potential, and measuring the current passing through the system to obtain a current-frequency relation curve of the sensing device; according to the relation between the current, the capacitance reactance and the frequency, extending the linear part of the obtained current-frequency relation curve, and obtaining the capacitance of the system through the intercept, wherein the capacitance of the system is the serial capacitance of the intrinsic capacitance and the interface capacitance of the conductive composite material;
under the condition that the external alternating current potential is unchanged, continuously changing the magnitude of the external direct current potential, and measuring the current passing through the system to obtain a current-grid voltage relation curve of the sensing device; and converting the current-gate voltage relation curve into a capacitance-gate voltage relation curve according to the relation between the current and the capacitance and the relation between the capacitance and the capacitance.
The evaluation of the liquid environment in which the sensing device is located according to the test result specifically comprises:
drawing capacitance-voltage relation curves measured under different liquid environments on the same coordinate system, and judging the liquid environments according to the regular change of the curves along with the occurrence of the molecular concentration in the solution;
obtaining the intrinsic capacitance of the system under the same external direct current potential under different liquid environments, and judging the liquid environments according to the regular change of the intrinsic capacitance along with the occurrence of the molecular concentration in the solution;
and obtaining the minimum value, inflection point and turning point bias voltage values of the capacitance-grid voltage relation curves under different liquid environments, and judging the liquid environment according to the regular change of the bias voltage values along with the occurrence of the molecular concentration in the solution.
The conductive composite material is formed by compounding a conductive two-dimensional material and a stable matrix material, wherein the weight percentage of the conductive two-dimensional material is 0-99.9%;
the conductive two-dimensional material comprises graphene and derivatives thereof, single-layer or multi-layer transition metal sulfide, perovskite with a two-dimensional nano structure, graphite alkyne and metal nano sheets;
the stable matrix material comprises oxide ceramics of silicon dioxide, aluminum dioxide, titanium dioxide, zirconium dioxide and hafnium dioxide, nitride ceramics of silicon nitride and boron nitride, carbide ceramics of silicon carbide, metal ceramics, functional ceramics and ceramic composites thereof, polyaniline, epoxy resin, phenolic resin, PBS, PVC organic matters and composites thereof.
The preparation method of the conductive composite material comprises the following steps:
uniformly dispersing the conductive two-dimensional material in a stable substrate material on an atomic scale by adopting an intercalation method, an ultrasonic treatment dispersion method or a powder mixing method, and further promoting and inducing the two-dimensional material sheet layer to realize directional arrangement in a ceramic substrate by adopting a pressure sintering method and an atmosphere pressure sintering method; the edges of the two-dimensional material are obtained by cutting or dissociating in a direction perpendicular to the oriented alignment of the two-dimensional material.
The two-dimensional material edge is further optimized by adding grinding, chemical etching and physical etching after the edge is obtained by cutting or dissociating in the direction perpendicular to the orientation arrangement of the two-dimensional material.
The surface chemical modification is carried out after the edges of the two-dimensional material are obtained by cutting or dissociating in the direction perpendicular to the orientation arrangement of the two-dimensional material, so as to modify the receptor specifically bound with the specific target substance.
The preparation method of the sensing device comprises the following steps:
coating an adhesive on the stable substrate, fixing the conductive composite material, and exposing a cutting surface for sensing as an upper surface; coating conductive adhesive on one side of the conductive composite material to be led out as an electrode; the non-sensing portion is encapsulated with an insulating material to create a liquid channel that exposes only the sensing surface and prevents liquid from contacting the electrodes.
The invention has the beneficial effects that:
1. the invention has the advantages of low cost, convenient preparation, controllable structure, high stability, good biocompatibility and repeated use;
2. the composite material can be repeatedly used for molecular detection, so that the service life of the sensor is greatly prolonged, the cost is reduced, and the operation is simple;
3. the optical and mechanical properties of the graphene can be utilized to realize photoelectric coupling and flexible sensor design, and the graphene quantum capacitor and a proper inductor form an LC oscillating circuit, so that the sensor is read in a wireless mode, and the instant sensing and information communication of the Internet can be realized;
4. the invention utilizes the local field enhancement effect of the exposed graphene edge and has extremely high detection sensitivity.
Drawings
FIGS. 1 (a) (b) (c) (d) are each a sensor device fabrication process;
FIG. 2 is an SEM image of a cross section of a graphene composite;
FIG. 3 is a field simulation diagram of a conductive composite;
fig. 4 (a) (b) is a diagram of an actual building circuit and an equivalent circuit of the test system, respectively;
FIG. 5 is a graph of intrinsic capacitance measurement current versus frequency;
FIG. 6 is a graph of capacitance-voltage curve measurement;
FIG. 7 is a graph of capacitance-gate voltage relationship;
FIG. 8 is a graph showing the time dependence of capacitance for different pH values.
Detailed Description
The invention provides a biochemical molecule detection method based on a quantum capacitance effect of a conductive composite material, and the invention is further described below with reference to the accompanying drawings and specific embodiments.
Sensor structure:
the sensing device mainly comprises a conductive composite material serving as a sensing element, an electrode led out and a liquid tank obtained by encapsulation.
The conductive composite material can be obtained by compounding a two-dimensional conductive material with a quantum capacitance effect with a stable matrix material. The size is not required. For ease of handling, 1 to 50mm is preferred.
The two-dimensional conductive material has a limited energy state density and thus an intrinsic capacitance is extremely small. Suitable two-dimensional conductive materials include, but are not limited to, graphene and its derivatives, single or multi-layer transition metal sulfides, two-dimensional nanostructured perovskites, graphite alkynes, metal nanoplatelets, and the like.
Generally, the matrix material is inexpensive, stable, and easy to prepare. The base material includes, but is not limited to, oxide ceramics such as silica, alumina, titania, zirconia, and hafnia, nitride ceramics such as silicon nitride and boron nitride, carbide ceramics such as silicon carbide, various cermets, functional ceramics, and ceramic composites thereof, and organic matters such as polyaniline, epoxy resin, phenolic resin, PBS, PVC, and the like.
The surface of the conductive composite material can be obtained by cutting or dissociating, the edge of the conductive composite material can be obtained by cutting or dissociating in the direction vertical to the directional arrangement of the two-dimensional material, and the treatment such as grinding, chemical etching, physical etching and the like can be added or not added to further optimize the edge of the two-dimensional material, and the surface chemical modification can be carried out or not, so that the receptor specifically combined with the specific target substance can be modified.
The extraction electrode includes, but is not limited to, a conductor material such as gold, silver, copper, etc. In a preferred embodiment, the extraction electrode is silver.
Materials for the liquid tank include, but are not limited to, insulators such as non-toxic epoxy, silicone, and the like. In a preferred embodiment, the material encapsulating the liquid tank is a biocompatible silicone.
The detection method comprises the following steps:
quantum capacitance testing methods include, but are not limited to, current-frequency curve measurement and CV curve measurement of the sensing device.
The frequency range of the applied varying alternating potential may be 1mHz to 10mHz when the current-frequency curve measurement is performed. The amplitude of the applied ac potential is typically in the form of a sine wave, which may be from 1mV to 100mV, preferably 7.07mV. The DC potential may be set to-10V to 10V, and in a preferred embodiment 0V.
In performing the CV curve measurement, the frequency range of the applied constant ac potential may be determined by judging the linear portion of the current-frequency relationship curve, or the like, and the amplitude thereof may be 1mV to 100mV, preferably 7.07mV. The applied DC potential may vary from-10V to 10V, and in a preferred embodiment from-1V to 1V. The current-gate voltage dependence is converted into a capacitance-gate voltage dependence according to the current-capacitance relationship, i.e., c=i/2pi fV.
Detection of biochemical molecules:
in one embodiment, the method is a method for sensing biochemical molecules. In this embodiment, the working electrode is in contact with a carrier medium that may include the molecule, and the conductive composite is responsive to the presence of the molecule to the quantum capacitance property of the applied potential, a particular quantum capacitance property will be obtained if the carrier medium does contain the molecule. Similarly, as the concentration of molecules in the carrier medium changes, so does the quantum capacitance properties.
The carrier medium is preferably in liquid form, although it may also be a gaseous medium. The carrier liquid (or gas carrier) may be any liquid (or gas) that can suspend or dissolve (or disperse) the substance. In one embodiment, the carrier comprises a pH buffered solution.
The sensing device includes a receptor moiety capable of binding the substance, and the sensing device is sensitive to the binding of the substance to the receptor moiety in response to a quantum capacitance response to an applied potential. Preferably, the receptor moiety is capable of specific binding of the substance. By "capable of specifically binding to the substance" is generally meant that the binding constant for the substance is at least 50 times, preferably at least 100 times, more preferably at least 200 times greater than the binding constant for any other substance present in the carrier medium.
Examples of receptor moieties include antibodies, antibody fragments, nucleic acids, nucleic acid aptamers, oligosaccharides, peptides, and proteins. Preferably, the receptor moiety is selected from antibodies, nucleic acids and peptides. The most preferred receptor moiety is an antibody.
The antibody or antibody fragment may be selected from one or more of the IgA, igD, igE, igG and IgM classes. The antibody selectively binds to a substance of interest. The antibody or antibody fragment may be derived from a mammal, including but not limited to, a mammal selected from the group consisting of human, mouse, rat, rabbit, goat, sheep, and horse. The nucleic acid aptamer may be selected from peptide aptamers, DNA aptamers, and RNA aptamers.
Obviously, the choice of acceptor moiety for a given electrode depends on the identity of the substance of interest (i.e., the "target" of interest). For example, the target may be an alpha synuclein (alpha-sync), in which case the receptor moiety typically comprises or consists of an anti-alpha-synuclein. The target species that the user wishes to detect/sense may or may not be present in the carrier medium, optionally together with one or more other non-target species. Most commonly, the method is a method for determining the concentration of the target species in the carrier medium.
Examples of target species include those selected from one or more markers of CRP protein, insulin, neurodegeneration, cancer, myocardial infarction, diabetes, and general trauma.
More generally, suitable target species for detection according to the methods of the invention include proteins, polypeptides, antibodies, nanoparticles, drugs, toxins, hazardous gases, hazardous chemicals, explosives, viral particles, cells, multicellular organisms, cytokines and chemokines, gametophytes, organelles, lipids, nucleic acid sequences, oligosaccharides, chemical intermediates of metabolic pathways, and macromolecules. In a preferred embodiment, the target species comprises, consists essentially of, or consists entirely of a biochemical molecule, more suitably a biological macromolecule, most suitably a polypeptide. Biomarkers are one example of biochemical molecules of particular interest.
Environmental parameter sensing:
the method can be used for sensing the change of environmental parameters of the environment where the electrode is located. Examples of such environmental parameters include the temperature of the environment, the light intensity of the environment (e.g., the intensity of visible light, or alternatively or additionally the intensity of ultraviolet light), and the humidity of the environment. In such methods, light, temperature or environmental/surface water interactions affect the measured quantum capacitance properties of the sensing device.
FIGS. 1 (a) (b) (c) (d) are each a sensor device fabrication process; the sensor device is prepared according to the following steps:
(a) Coating a biocompatible silica gel on the stable substrate as a binder;
(b) Fixing the conductive composite material on the adhesive: the reduced graphene oxide/silicon dioxide ceramic composite material has the reduced graphene oxide content of 5%;
(c) Coating silver colloid on one side of the composite material to be led out as an electrode;
(d) And packaging the non-sensing part by using silica gel to manufacture a liquid groove so as to prevent the liquid from contacting with the electrode.
FIG. 2 is an SEM image of a cross section of a graphene composite; the conductive composite used was characterized using a scanning electron microscope: the graphene oxide (5%) and silica ceramic composite material was reduced, and the characterization result is shown in fig. 2. The edges of the conductive graphene are clearly visible, and the graphene is dispersed on a fuzzy non-conductive ceramic matrix in a lamellar manner, so that the reduced graphene oxide is proved to be uniformly dispersed in the ceramic matrix in an atomic scale by adopting an intercalation method.
FIG. 3 is a field simulation diagram of a conductive composite; the field simulation was performed on the edge of reduced graphene oxide using comsol, and the results are shown in fig. 3. A sharp voltage drop is observed near the graphene edge, resulting in a prominent local electric field enhancement at the graphene edge, which is an order of magnitude stronger than just 1nm from the edge.
Example one Quantum capacitance test
Reagent: standard hydrochloric acid solution and sodium hydroxide medicine of 0.1mol/L
The device comprises: MFLI lock-in amplifier, probe station, saturated calomel electrode
The experimental method comprises the following steps:
a) Serial concentration hydrochloric acid and sodium hydroxide solution (pH=3-11) preparation
9.9mL of deionized water was taken out by a pipette, and 100. Mu.L of a standard hydrochloric acid solution of 0.1mol/L was added dropwise to the deionized water by the pipette to obtain a 1mM hydrochloric acid solution (pH=3). The hydrochloric acid solution was diluted ten times with deionized water to obtain a hydrochloric acid solution having ph=4. After that, a ten-fold dilution method was also adopted to obtain hydrochloric acid solutions of ph=5, 6.
0.4g of sodium hydroxide medicine is weighed by an electronic balance and dissolved in 10mL of deionized water to obtain 1mol/L sodium hydroxide solution. 9.09mL of deionized water was taken with a pipette, and 10. Mu.L of a 1mol/L standard hydrochloric acid solution was added dropwise to the deionized water with the pipette to obtain a 1mM sodium hydroxide solution (pH=11). Ten times the dilution of this solution with deionized water gave sodium hydroxide solution at ph=10. After that, a ten-fold dilution method was also employed to obtain sodium hydroxide solutions of ph=5, 6.
b) Test system circuit construction
Fig. 4 (a) (b) is a diagram of an actual building circuit and an equivalent circuit of the test system, respectively; the test was performed with a three electrode system. The graphene/ceramic composite material is used as a working electrode, a calomel electrode is used as a reference electrode, and an inert graphite electrode is used as a counter electrode. And (3) dripping the solution into the liquid tank to enable the working electrode and the reference electrode to be in contact with the solution, thereby forming a circuit.
c) Intrinsic capacitance measurement
In the circuit shown in FIG. 4After the solution is dripped into the device solution tank, the frequency of the alternating voltage output by the power supply is changed, and the current in the circuit is measured. During the test, the AC voltage was set to 7.07mV (i.e., V pk =10mv), the dc voltage is set to 0V, the frequency variation range is 1-100kHz, and one sweep is performed back and forth; the frequency sampling points are selected in a log mode, 101 sampling points are obtained through one test, and a current-frequency relation curve is shown in fig. 5. The intercept (i.e. 2pi fC, where total capacitance in series c=c) can be obtained by linear fitting based on an RC equivalent circuit Q C DL /(C Q +C DL ) And then calculate the quantum capacitance C of the graphene/ceramic composite material Q (C DL Typically a known constant). The linear portion of fig. 5 is fitted with a linear function to obtain the intercept, and the intrinsic capacitance c=1 μf/cm of the material is calculated to obtain 2 。
d) Capacitance-gate voltage relationship curve determination
In the circuit shown in FIG. 4, after the prepared solution is dropped into the device solution tank, the DC voltage (i.e., solution gate voltage V) output from the power supply is changed ref ) And measuring the current in the circuit, and calculating according to C=I/2pi Vf to obtain a capacitance-gate voltage relation curve. During the test, the AC voltage was set to 7.07mV (i.e., V pk =10mv), 77.77Hz, the dc voltage setting ranges from-1V to 1V, and scanning is performed back and forth, and sampling is performed every 0.02V, resulting in a capacitance-voltage relationship curve as shown in fig. 6. As shown in fig. 6, the capacitance-voltage curve exhibits a V-shape, exhibiting good field effect performance. In practical testing, it is not excluded to obtain a capacitance-voltage curve of inverted V-shape or other shape.
Examples sensory response test of Potassium dichloride concentration
Reagent: 1mol/L potassium chloride solution
The device comprises: MFLI lock-in amplifier, probe station, silver/silver chloride electrode
The experimental method comprises the following steps:
a) Preparation of potassium chloride solution series
9mL of deionized water is taken by a liquid-transfering gun, and 1mL of 1mol/L potassium chloride solution is taken by the liquid-transfering gun and dripped into the deionized water, so that 100mM potassium chloride solution is obtained. The ten-fold dilution method was repeated to obtain potassium chloride solutions having concentrations of 10mM and 1mM, respectively.
b) Test system circuit construction
The test was performed with a three electrode system. The graphene/titanium dioxide material is used as a working electrode, a reference electrode is a silver/silver chloride electrode, and a counter electrode is an inert graphite electrode. And (3) dripping the solution into the liquid tank to enable the working electrode and the reference electrode to be in contact with the solution, thereby forming a circuit.
c) Capacitance curve determination of different potassium chloride concentrations
After the prepared potassium chloride solution with different concentrations is dripped into the solution tank of the device, the direct current voltage output by the power supply (namely the solution grid voltage V ref ) And measuring the current in the circuit, and calculating according to C=I/2pi Vf to obtain a capacitance-gate voltage relation curve. During testing, the AC voltage was set to 0.707mV (i.e., V pk =1 mV), 121Hz, the dc voltage setting ranges from-0.5V to 0.5V, sampling is performed every 0.01V, and the capacitance-voltage relationship is shown in fig. 7.
As shown in fig. 7, the capacitance-voltage curve exhibits a Λ shape, exhibiting a shape different from that of the conventional graphene capacitance curve; additionally, upon increasing the potassium chloride concentration, the "Λ" point is shifted forward (< 5mV dec-1). This smaller magnitude of inductive reaction compared to the Nernst limit of 59.2mV dec-1 at room temperature indicates that the graphene edges adsorb negatively charged Cl-ions relatively weakly. That is, both K+ and Cl-ions are chemically inert, with less tendency to adsorb specifically at the edges of graphene. Thus, there is no significant ion charge doping effect.
Example three pH sensory response test
Reagent: buffer solution with ph=3-12
The device comprises: MFLI lock-in amplifier, probe station, silver/silver chloride electrode
The experimental method comprises the following steps:
a) Test system circuit construction
The test was performed with a three electrode system. The graphene/silicon nitride material is used as a working electrode, a reference electrode is a silver/silver chloride electrode, and a counter electrode is an inert graphite electrode. And (3) dripping the solution into the liquid tank to enable the working electrode and the reference electrode to be in contact with the solution, thereby forming a circuit.
b) Capacitance time dependent curve determination of different pH values
And dripping the prepared buffer solution into a device solution tank, changing the buffer solution every ten minutes, changing the pH value from low to high, and measuring the capacitance in the circuit. During the test, the AC voltage was set to 7.07mV (i.e., V pk =10mv), the dc voltage was set to 0V, the frequency was 216Hz, and a time-dependent curve of the capacitance change amount was calculated from the magnitude of the initial capacitance. The capacitance time dependence curves for different pH values are shown in FIG. 8. The capacitance changes obviously along with the change of the concentration of hydrogen ions, and the sensing performance is good.
In summary, the invention utilizes the graphene composite material to carry out biochemical molecular detection based on quantum capacitance effect, and the graphene composite material shows a field effect that the capacitance can be regulated and controlled by the gate voltage through the measurement of the intrinsic capacitance and the measurement and investigation of the capacitance-voltage relation curve, and has small intrinsic capacitance, easy detection and high detection sensitivity.
Claims (10)
1. The biochemical molecule detection method based on the quantum capacitance effect of the conductive composite material is characterized by comprising the following steps of:
testing by adopting a three-electrode system of a working electrode, a reference electrode and a counter electrode, and enabling the working electrode and the reference electrode to be in contact with the solution to form a loop by dripping the solution into a liquid tank;
and carrying out quantum capacitance test by using a lock-in amplifier or an impedance analyzer, evaluating the liquid environment of the sensing device according to the test result, and detecting the concentration of biochemical molecules in the liquid.
2. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to claim 1, wherein the working electrode is conductive composite material, the reference electrode is calomel or silver chloride, and the counter electrode is graphite or platinum electrode.
3. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to claim 1, wherein the biochemical molecules comprise hydrogen ions, hydroxyl ions, potassium ions, chloride ions, DNA, RNA, glucose.
4. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to claim 1, wherein the quantum capacitance test using a lock-in amplifier or an impedance analyzer is specifically as follows:
under the condition that the external direct current potential is unchanged, continuously changing the frequency of the external alternating current potential, and measuring the current passing through the system to obtain a current-frequency relation curve of the sensing device; according to the relation between the current, the capacitance reactance and the frequency, extending the linear part of the obtained current-frequency relation curve, and obtaining the capacitance of the system through the intercept, wherein the capacitance of the system is the serial capacitance of the intrinsic capacitance and the interface capacitance of the conductive composite material;
under the condition that the external alternating current potential is unchanged, continuously changing the magnitude of the external direct current potential, and measuring the current passing through the system to obtain a current-grid voltage relation curve of the sensing device; and converting the current-gate voltage relation curve into a capacitance-gate voltage relation curve according to the relation between the current and the capacitance and the relation between the capacitance and the capacitance.
5. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to claim 1, wherein the step of evaluating the liquid environment of the sensing device according to the test result comprises:
drawing capacitance-voltage relation curves measured under different liquid environments on the same coordinate system, and judging the liquid environments according to the regular change of the curves along with the occurrence of the molecular concentration in the solution;
obtaining the intrinsic capacitance of the system under the same external direct current potential under different liquid environments, and judging the liquid environments according to the regular change of the intrinsic capacitance along with the occurrence of the molecular concentration in the solution;
and obtaining the minimum value, inflection point and turning point bias voltage values of the capacitance-grid voltage relation curves under different liquid environments, and judging the liquid environment according to the regular change of the bias voltage values along with the occurrence of the molecular concentration in the solution.
6. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to any one of claims 1 to 5, wherein the conductive composite material is formed by compounding conductive two-dimensional material with stable matrix material, wherein the weight percentage of the conductive two-dimensional material is 0 to 99.9%;
the conductive two-dimensional material comprises graphene and derivatives thereof, single-layer or multi-layer transition metal sulfide, perovskite with a two-dimensional nano structure, graphite alkyne and metal nano sheets;
the stable matrix material comprises oxide ceramics of silicon dioxide, aluminum dioxide, titanium dioxide, zirconium dioxide and hafnium dioxide, nitride ceramics of silicon nitride and boron nitride, carbide ceramics of silicon carbide, metal ceramics, functional ceramics and ceramic composites thereof, polyaniline, epoxy resin, phenolic resin, PBS, PVC organic matters and composites thereof.
7. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to claim 6, wherein the method for preparing the conductive composite material is as follows:
uniformly dispersing the conductive two-dimensional material in a stable substrate material on an atomic scale by adopting an intercalation method, an ultrasonic treatment dispersion method or a powder mixing method, and further promoting and inducing the two-dimensional material sheet layer to realize directional arrangement in a ceramic substrate by adopting a pressure sintering method and an atmosphere pressure sintering method; the edges of the two-dimensional material are obtained by cutting or dissociating in a direction perpendicular to the oriented alignment of the two-dimensional material.
8. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to claim 7, wherein the optimization of two-dimensional material edges is further realized by adding grinding, chemical etching and physical etching after cutting or dissociating in a direction perpendicular to the alignment of two-dimensional material to obtain the edges.
9. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to claim 7 or 8, wherein the receptor specifically bound to a specific target substance is modified by performing surface chemical modification after cleavage or dissociation in a direction perpendicular to the alignment of two-dimensional material to obtain its edge.
10. The method for detecting biochemical molecules based on quantum capacitance effect of conductive composite material according to claim 1 or 4, wherein the method for manufacturing the sensing device is as follows:
coating an adhesive on the stable substrate, fixing the conductive composite material, and exposing a cutting surface for sensing as an upper surface; coating conductive adhesive on one side of the conductive composite material to be led out as an electrode; the non-sensing portion is encapsulated with an insulating material to create a liquid channel that exposes only the sensing surface and prevents liquid from contacting the electrodes.
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