CN105758849B - A kind of preparation method and application of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure - Google Patents
A kind of preparation method and application of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure Download PDFInfo
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- CN105758849B CN105758849B CN201610148497.0A CN201610148497A CN105758849B CN 105758849 B CN105758849 B CN 105758849B CN 201610148497 A CN201610148497 A CN 201610148497A CN 105758849 B CN105758849 B CN 105758849B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y30/00—Nanotechnology for materials or surface science, e.g. nanocomposites
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
Abstract
The invention discloses a kind of preparation method of the biological immune sensing device for possessing electrogenerated chemiluminescence and the flourishing strategy of optical electro-chemistry dual signal based on function nano material structure, prepared sensor operations are simple, easy to carry, detection is fast, cost is low, quick, the Sensitive Detection to stilbestrol available for fields such as daily production and lifes.
Description
Technical field
The present invention relates to a kind of preparation method for being used to detect the sensor of stilbestrol, sensor integration electrochemistry
Luminous and optical electro-chemistry dual-use function.Belong to Nano-function thin films and electrochemical biosensor analysis technical field.
Background technology
After environmental estrogens refer to that one kind enters body, there is the synthesis, release, fortune of normal endocrine material in interfering bodies
The processes such as defeated, combination, metabolism, activation or the function of suppressing internal system, organism stability and regulation and control are maintained to make so as to destroy
Compound, environmental estrogens species is various, including artificial-synthetic compound and natural plant estrogen, is distributed widely in certainly
In right boundary.Diethylstilbestrol is a kind of artificial synthesized non steroidal estrogen material.At present, the method for detecting diethylstilbestrol is main
There are chromatography, mass spectrography etc..Such method instrument is valuable, complex operation, and laboratory personnel could be examined after needing professional training
Survey.Therefore, R＆D costs are low, detection is fast, the diethylstilbestrol sensor of high sensitivity, high specificity is significant.
Electrochemical biosensor analytical technology increasingly obtains the weight of people due to the advantage such as easy to operate, detection speed is fast
Depending on.The electrochemical biosensor analytical technology for being presently used for detecting diethylstilbestrol mainly has electrochemistry according to detection means to divide
Three kinds of sensor, electrochemical luminescence sensor and optical electro-chemistry sensor.Wherein, electrochemical luminescence sensor and optical electro-chemistry pass
Sensor has the characteristics that few background signal noise, high sensitivity, testing cost are low relative to electrochemical sensor, in recent years by
Increasing researcher is of interest.
Electrochemical luminescence is also referred to as electrogenerated chemiluminescence, and it is special to refer to produce some in electrode surface by electrochemical method
Material, form excitation state between these materials or by electron transmission between other components in system, returned by excitation state
Return to ground state and produce luminescence phenomenon.Electrochemical luminescence sensor is the decorative material by changing electrode surface, with analyzing produce
Raw electrochemical luminescence, in optimal conditions, realized according to the associated change of analyte concentration and electrochemical luminescence intensity to analysis
The qualitative and quantitative analysis of thing.
Optical electro-chemistry sensor is to cause electron-hole pair to be separated based on additional light source activation Electrophotosensitivmaterial material,
Under suitable potential condition partially, quick transmission of the electronics on electrode, semiconductor and trim and analyte is realized, and formed
Photoelectric current.In optimal conditions, the change of analyte concentration can directly affect the size of photoelectric current, can be according to the change of photoelectric current
Change the qualitative and quantitative analysis realized to analyte.
But because electrochemical luminescence sensor needs external optical signal to catch equipment such as photodiode etc., and photoelectricity
Chemical sensor needs peripheral hardware light source to excite Electrophotosensitivmaterial material, and this have impact on the convenient of the two operation to a certain extent
Property, limit them and be more widely applied in actual production, life.Therefore, design, prepare more simple, fast inspection
The electrochemical biosensor analytical technology for surveying diethylstilbestrol has highly important practical value.
The content of the invention
It is an object of the invention to provide a kind of simple to operate, easy to carry, detection is fast, cost is low diethylstilbestrol without enzyme
The preparation method of sensor, prepared sensor, available for the fields such as daily production and life to diethylstilbestrol it is quick,
Sensitive Detection.Based on this purpose, the present invention is in same electrolytic cell, using four electrode systems, i.e. two working electrodes, one it is right
Electrode and a reference electrode, wherein working electrode 1 use the molybdenum disulfide composite Zn-MoS of zinc doping2Gold-supported@platinum
Nanometer rods Au@Pt and electropolymerization luminol are modified jointly, and as electrochemical luminescence working electrode W1, working electrode 2 uses
Reduced graphene carried titanium dioxide nano-film sol rGO/TiO2Modified jointly with diethylstilbestrol antibody, as photoelectrochemical
Learn working electrode W2.When being detected, after adding the hydrogen peroxide of fixed concentration in electrolytic cell, apply step electricity on W1
Pressure, due to Zn-MoS2The work of catalyst activity preferably can be loaded and kept with bigger serface and good electric conductivity
With, and Au@Pt have the characteristics of more avtive spots compared to simple platinum catalyst so that luminol more fills with hydrogen peroxide
Divide, more effectively reacted, produce the electrochemical luminescence of sufficient intensity, this is just equivalent to " turning on light ", when step voltage is 0
When, electrochemical luminescence disappears, and this just equivalent to " turning off the light ", at the same time applies constant voltage, due to rGO/ always on W2
TiO2It can be marked because lighting to excite caused by electrochemical luminescence causes electron-hole pair to be separated in stilbestrol secondary antibody
On hemin Hemin catalyzing hydrogen peroxides produce oxygen, hydrogen peroxide is turned into hole " donor ", so as on W2 must
To photoelectric current, when electrogenerated chemiluminescence disappears, i.e., when " turning off the light ", photoelectric current disappears immediately.Due in fixed concentration of hydrogen peroxide
On the premise of, photoelectric current size and Hemin concentration positive correlations, when stilbestrol concentration is bigger in measured object, when with an anti-immunity
When being attached on W2, then immune binding marker has the concentration of Hemin stilbestrol secondary antibody will be bigger, caused photoelectric current
It is bigger, therefore by recording the size i.e. achievable detection to stilbestrol of photoelectric current.
Based on above inventive principle, the concrete technical scheme that the present invention uses is as follows：
1. a kind of preparation method of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure, it is special
Point is that preparation process is：
（1）Electrochemical luminescence working electrode W1 preparation method, described W1 are by Zn-MoS2Load Au@Pt and voltolisation
The ITO electro-conductive glass that luminol is modified jointly is closed, is characterized in, specific preparation process is：
1）Using ITO electro-conductive glass as working electrode, in electrode surface drop coating Zn-MoS2, area coverage is 1 cm × 1
Cm, dry at room temperature；
2）By 1）Obtained working electrode, in Zn-MoS2Surface drop coating Au@Pt, area coverage are the cm of 1 cm × 1, room
Dried under temperature；
3）By 2）Obtained working electrode, immerse in electrolyte, immersion area is Zn-MoS2Covered after load Au@Pt
Area, electrochemical deposition is carried out to working electrode using three-electrode system, takes out working electrode after deposition, it is clear using ultra-pure water
Wash, lucifuge is dried at 4 DEG C, and electrochemical luminescence working electrode W1 is made；
Described Zn-MoS2For the molybdenum disulfide two-dimension nano materials doped with zinc ion, described two-dimension nano materials
For scraps of paper shape, thickness is about 10 ~ 20 nm；Described Zn-MoS2Preparation method be：By 0.005 ~ 0.01 g zinc chloride ZnCl2、
2 ~ 6 mL 0.1 mol/L ascorbic acid solutions, 0.5 ~ 1.5 mL sodium molybdate Na2MoO4Solution and 0.01 ~ 0.03 g vulcanizations
Sodium Na2S, after being mixed and stirred for 15 minutes, it is put into reactor, at 150 ~ 220 DEG C, reacts 12 ~ 16 hours；It is cooled to room temperature
Afterwards, using deionized water centrifuge washing, it is dried in vacuo at 40 DEG C, that is, Zn-MoS is made2, it is dissolved in deionized water
Zn-MoS is made2Solution；
Described Au@Pt are the rod-like nano material of core shell structure, and the rod-like nano material of the core shell structure is with gold
Nanometer rods are core, and nano platinum particle is the rod-like nano material of shell, and the gold nanorods are bar-like golden nanometer particle, and length is
20~40 nm；
Described electrolyte is the sulfuric acid solution containing luminol, and the concentration of luminol is 1 ~ 10 in described electrolyte
Mmol/L, sulfuric acid concentration are 0.1 ~ 1.0 mol/L；
Described three-electrode system, including working electrode, reference electrode and to electrode, described reference electrode is that saturation is sweet
Mercury electrode, described is platinum electrode to electrode；
Described electrochemical deposition process, for the electrochemical method used for cyclic voltammetry, starting voltage is -0.2 V, eventually
Only voltage is 1.5 V, and it is 100 mv/s to sweep speed, the circle of circulation 20 ~ 30；
（2）Optical electro-chemistry working electrode W2 preparation method, described W2 are by rGO/TiO2It is total to stilbestrol antibody
With the ITO electro-conductive glass of modification, it is characterized in, specific preparation process is：
1）Using ITO electro-conductive glass as working electrode, in the μ L rGO/TiO of electrode surface drop coating 8 ~ 122, dry at room temperature；
2）By 1）In obtained working electrode be put into Muffle furnace, made annealing treatment at 450 DEG C, cooled down after processing
To room temperature；
3）By 2）In obtained μ L stilbestrol antibody-solutions of working electrode surface drop coating 8 ~ 12, dry at 4 DEG C, do
Cleaned with ultra-pure water after dry, dried at 4 DEG C, optical electro-chemistry working electrode W2 is made；
Described rGO/TiO2For the aqueous solution of reduced graphene carried titanium dioxide nanometer sheet, the nano titania
Piece is the titanium dioxide nano-particle of square sheet, and the length of side is 60 ~ 80 nm；
The concentration of described stilbestrol antibody-solutions is 300 μ g/mL；
（3）Preparation method of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure：
1）By in the relative insertion electrolytic cell of W1 and W2 conductive one side, W1 and W2 spacing are the cm of 0.5 cm ~ 1.5；
2）It is to electrode CE by reference electrode RE, platinum electrode of Ag/AgCl, inserts in electrolytic cell, it is common with W1 and W2
Form four electrode systems；
3）It is 1 mmol/L's that NaOH solution that 10 mL pH value are 11 ~ 13 and 0.2 mL concentration are added in electrolytic cell
Hydrogenperoxide steam generator；
4）By 1）~3）Obtained four electrode systems and electrolytic cell are placed in magazine, that is, are made and are based on Two-dimensional Composites
The difunctional stilbestrol of structure is without enzyme sensor.
2. the difunctional stilbestrol of the present invention based on Two-dimensional Composites structure is applied to without enzyme sensor
The detection of diethylstilbestrol, it is characterized in that specific detection method is：
（1）Be added dropwise the stilbestrol standard liquid of 10 μ L various concentrations on W2, after hatching 30 min, stilbestrol with
Stilbestrol antibody on W2 carries out immune combination, after flushing, then be added dropwise 10 μ L hemins Hemin marks alkene it is female
Phenol secondary antibody, after hatching 30 min, stilbestrol secondary antibody and the stilbestrol of Hemin marks carry out immune combination, after flushing, are made
W2 to be measured；
（2）Using electrochemical workstation, step voltage, initial voltage are applied to W1 using the method for step voltage on W1
For 0 v, step voltage is 0.7 ~ 0.9 v, and snap time is 10 ~ 30 s；Meanwhile the m- current methods when being used on W2 to be measured
Constant voltage is applied to W2 to be measured, voltage is 0 ~ 0.6 v；Electric current on W2 to be measured can with the increase of stilbestrol concentration phase
It should increase, according to the relation between gained electric current increment value and stilbestrol concentration, drawing curve；；
（3）Stilbestrol solution to be measured is replaced to the standard liquid of stilbestrol, according to（1）With（2）Described alkene is female
Phenol detection method is detected, and show that stilbestrol to be measured is molten with the working curve drawn according to resulting electric current increment value
The concentration of liquid；
The concentration of the stilbestrol secondary antibody of described Hemin marks is 300 μ g/mL.
The useful achievement of the present invention
（1）Difunctional stilbestrol of the present invention based on Two-dimensional Composites structure prepares letter without enzyme sensor
It is single, it is easy to operate, without external accessory, using the miniaturization of detection device, portability, and realize to diethylstilbestrol
Quickly, sensitive, high selectivity detection, has wide market development prospect；
（2）The present invention realizes electrochemistry first in same electrolytic cell using four electrode systems detection stilbestrol
Luminous signal amplification strategy difunctional with optical electro-chemistry.With the increase of stilbestrol concentration in electrolytic cell, modify on W2
Hemin mark stilbestrol secondary antibody will increase, also imply that Hemin to hydrogen peroxide catalyzed raising.Such one
Come, on the one hand, so that electrochemical luminescence intensity is linearly increasing, the photoelectricity cleanliness increase excited；On the other hand, hydrogen peroxide
As electron donor so that photoelectricity cleanliness increases in optical electro-chemistry reaction.Therefore, two kinds of sides of electrochemical luminescence and optical electro-chemistry
Method is reacted, interacted jointly in same electrolytic cell, under same electrochemical workstation, is realized and is detected telecommunications to stilbestrol
Number dual amplification, detection sensitivity and detection limit are drastically increased, simultaneously because do not use biology enzyme, so as to detection
Environmental requirement is more loose, has important scientific meaning and market application value.
Embodiment
A kind of difunctional stilbestrol based on Two-dimensional Composites structure of embodiment 1 is without enzyme sensor, specific system
It is for step：
（1）Zn-MoS2Preparation：
By 0.005 g zinc chloride ZnCl2, 2 mL 0.1 mol/L ascorbic acid solutions, 0.5 mL sodium molybdate Na2MoO4
Solution and 0.01 g vulcanized sodium Na2S, after being mixed and stirred for 15 minutes, it is put into reactor, at 150 DEG C, reacts 16 hours；
After being cooled to room temperature, using deionized water centrifuge washing, it is dried in vacuo at 40 DEG C, that is, Zn-MoS is made2, its is molten
Zn-MoS is made in deionized water2Solution.
（2）Electrochemical luminescence working electrode W1 preparation：
1）Using ITO electro-conductive glass as working electrode, in electrode surface drop coating（1）The Zn-MoS of middle preparation2Solution, coverage rate
Product is the cm of 1 cm × 1, is dried at room temperature；
2）By 1）Obtained working electrode, in Zn-MoS2Surface drop coating Au@Pt, area coverage are the cm of 1 cm × 1, room
Dried under temperature；
3）By 2）Obtained working electrode, immerse in electrolyte, immersion area is Zn-MoS2Covered after load Au@Pt
Area, electrochemical deposition is carried out to working electrode using three-electrode system, takes out working electrode after deposition, it is clear using ultra-pure water
Wash, lucifuge is dried at 4 DEG C, and electrochemical luminescence working electrode W1 is made；
Described Au@Pt are the rod-like nano material of core shell structure, and the rod-like nano material of the core shell structure is with gold
Nanometer rods are core, and nano platinum particle is the rod-like nano material of shell, and the gold nanorods are bar-like golden nanometer particle, and length is
20 nm；
Described electrolyte is the sulfuric acid solution containing luminol, and the concentration of luminol is 1 in described electrolyte
Mmol/L, sulfuric acid concentration are 0.1 mol/L；
Described three-electrode system, including working electrode, reference electrode and to electrode, described reference electrode is that saturation is sweet
Mercury electrode, described is platinum electrode to electrode；
Described electrochemical deposition process, for the electrochemical method used for cyclic voltammetry, starting voltage is -0.2V, eventually
Only voltage is 1.5V, and it is 100mv/s to sweep speed, the circle of circulation 20.
（3）Optical electro-chemistry working electrode W2 preparation：
1）Using ITO electro-conductive glass as working electrode, in the μ L rGO/TiO of electrode surface drop coating 82, dry at room temperature；
2）By 1）In obtained working electrode be put into Muffle furnace, made annealing treatment at 450 DEG C, cooled down after processing
To room temperature；
3）By 2）In obtained μ L stilbestrol antibody-solutions of working electrode surface drop coating 8, dry at 4 DEG C, after drying
Cleaned with ultra-pure water, dried at 4 DEG C, optical electro-chemistry working electrode W2 is made；
Described rGO/TiO2For the aqueous solution of reduced graphene carried titanium dioxide nanometer sheet, the nano titania
Piece is the titanium dioxide nano-particle of square sheet, and the length of side is 60 ~ 80 nm；
The concentration of described stilbestrol antibody-solutions is 300 μ g/mL；
（4）Preparation method of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure：
1）Will（2）The W1 of middle preparation and（3）The W2 of middle preparation is inserted in electrolytic cell face to face, and W1 is 0.5 with W2 spacing
cm；
2）It is to electrode CE by reference electrode RE, platinum electrode of Ag/AgCl, inserts in electrolytic cell, it is common with W1 and W2
Form four electrode systems；
3）The NaOH solution that 10 mL pH value are 11 and the peroxide that 0.2 mL concentration is 1 mmol/L are added in electrolytic cell
Change hydrogen solution；
4）By 1）~3）Obtained four electrode systems and electrolytic cell are placed in magazine, that is, are made and are based on Two-dimensional Composites
The difunctional stilbestrol of structure is without enzyme sensor.
A kind of difunctional stilbestrol based on Two-dimensional Composites structure of embodiment 2 is without enzyme sensor, specific system
It is for step：
（1）Zn-MoS2Preparation：
Preparation process is the same as Zn-MoS in embodiment 12The preparation method of solution, difference are： ZnCl2Addition be
0.008 g, the addition of ascorbic acid solution are 4 mL, Na2MoO4The addition of solution is 1.0 mL and Na2S addition is
0.02 g, at 220 DEG C, react 12 hours.
（2）Electrochemical luminescence working electrode W1 preparation：
With the preparation process of W1 in embodiment 1, difference is preparation process：Using in the present embodiment（1）Obtained
Zn-MoS2Modified electrode；Gold nanorods length is 30 nm；The concentration of luminol is 5 mmol/L in electrolyte, and sulfuric acid concentration is
0.5 mol/L, when cyclic voltammetry carries out electrochemical deposition, the circle of circulation 25.
（3）Optical electro-chemistry working electrode W2 preparation：
1）Using ITO electro-conductive glass as working electrode, in the μ L rGO/TiO of electrode surface drop coating 102, dry at room temperature；
2）By 1）In obtained working electrode be put into Muffle furnace, made annealing treatment at 450 DEG C, cooled down after processing
To room temperature；
3）By 2）In obtained μ L stilbestrol antibody-solutions of working electrode surface drop coating 10, dry at 4 DEG C, dry
Cleaned with ultra-pure water, dried at 4 DEG C afterwards, optical electro-chemistry working electrode W2 is made；
Remaining preparation process with W2 in embodiment 1.
（4）Preparation method of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure：
Preparation process is with embodiment 1, and difference is W1 and W2 spacing is 1.0 cm, and the NaOH added in electrolytic cell is molten
The pH value of liquid is 12.
A kind of difunctional stilbestrol based on Two-dimensional Composites structure of embodiment 3 is without enzyme sensor, specific system
It is for step：
（1）Zn-MoS2Preparation：
Preparation process is the same as Zn-MoS in embodiment 12The preparation method of solution, difference are： Zn Cl2Addition be
0.01 g, the addition of ascorbic acid solution are 6 mL, Na2MoO4The addition of solution is 1.5 mL and Na2S addition is
0.03 g, at 220 DEG C, react 12 hours.
（2）Electrochemical luminescence working electrode W1 preparation：
With the preparation process of W1 in embodiment 1, difference is preparation process：Using in the present embodiment（1）Obtained
Zn-MoS2Modified electrode；Gold nanorods length is 40 nm；The concentration of luminol is 10 mmol/L in electrolyte, sulfuric acid concentration
For 1.0 mol/L, when cyclic voltammetry carries out electrochemical deposition, the circle of circulation 30.
（3）Optical electro-chemistry working electrode W2 preparation：
1）Using ITO electro-conductive glass as working electrode, in the μ L rGO/TiO of electrode surface drop coating 122, dry at room temperature；
2）By 1）In obtained working electrode be put into Muffle furnace, made annealing treatment at 450 DEG C, cooled down after processing
To room temperature；
3）By 2）In obtained μ L stilbestrol antibody-solutions of working electrode surface drop coating 12, dry at 4 DEG C, dry
Cleaned with ultra-pure water, dried at 4 DEG C afterwards, optical electro-chemistry working electrode W2 is made；
Remaining preparation process with W2 in embodiment 1.
（4）Preparation method of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure：
Preparation process is with embodiment 1, and difference is W1 and W2 spacing is 1.5 cm, and the NaOH added in electrolytic cell is molten
The pH value of liquid is 13.
A kind of application of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure of embodiment 4
Difunctional stilbestrol based on Two-dimensional Composites structure prepared by embodiment 1 is applied to hexene without enzyme sensor
The detection of female phenol, its detecting step are：
（1）Be added dropwise the stilbestrol standard liquid of 10 μ L various concentrations on W2, after hatching 30 min, stilbestrol with
Stilbestrol antibody on W2 carries out immune combination, after flushing, then the stilbestrol secondary antibody of 10 μ L Hemin marks is added dropwise, incubates
After changing 30 min, stilbestrol secondary antibody and the stilbestrol of Hemin marks carry out immune combination, and after flushing, W2 to be measured is made；
（2）Using electrochemical workstation, step voltage, initial voltage are applied to W1 using the method for step voltage on W1
For 0 v, step voltage is 0.7 v, and snap time is 10 s；Meanwhile on W2 to be measured use when m- current methods to W2 to be measured
Apply constant voltage, voltage is 0 v；Electric current on W2 to be measured can accordingly increase with the increase of stilbestrol concentration, according to
Relation between gained electric current increment value and stilbestrol concentration, drawing curve；
（3）Stilbestrol solution to be measured is replaced to the standard liquid of stilbestrol, according to（1）With（2）Described alkene is female
Phenol detection method is detected, and show that stilbestrol to be measured is molten with the working curve drawn according to resulting electric current increment value
The concentration of liquid；
The concentration of the stilbestrol secondary antibody of described Hemin marks is 300 μ g/mL.
A kind of application of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure of embodiment 5
Difunctional stilbestrol based on Two-dimensional Composites structure prepared by embodiment 2 is applied to hexene without enzyme sensor
The detection of female phenol, its detecting step is in addition to following steps, and remaining step is the same as embodiment 4：
Step（2）Using electrochemical workstation, step voltage is applied to W1 using the method for step voltage on W1, initially
Voltage is 0 v, and step voltage is 0.8 v, and snap time is 20 s；Meanwhile m- current methods are treated when being used on W2 to be measured
Survey W2 and apply constant voltage, voltage is 0.3 v；Electric current on W2 to be measured can accordingly increase with the increase of stilbestrol concentration
Greatly, according to the relation between gained electric current increment value and stilbestrol concentration, drawing curve.
A kind of application of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure of embodiment 6
Difunctional stilbestrol based on Two-dimensional Composites structure prepared by embodiment 3 is applied to hexene without enzyme sensor
The detection of female phenol, its detecting step is in addition to following steps, and remaining step is the same as embodiment 4：
Step（2）Using electrochemical workstation, step voltage is applied to W1 using the method for step voltage on W1, initially
Voltage is 0 v, and step voltage is 0.9 v, and snap time is 30 s；Meanwhile m- current methods are treated when being used on W2 to be measured
Survey W2 and apply constant voltage, voltage is 0.6 v；Electric current on W2 to be measured can accordingly increase with the increase of stilbestrol concentration
Greatly, according to the relation between gained electric current increment value and stilbestrol concentration, drawing curve.
The difunctional stilbestrol based on Two-dimensional Composites structure prepared by the embodiment 1-3 of embodiment 7 passes without enzyme
Sensor, it is applied to the detection of stilbestrol according to embodiment 4-6 detecting step, there are excellent Detection results, detection is limited to
22 pmol/L。
Claims (6)
1. a kind of preparation method of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure, its feature exist
In using the molybdenum disulfide composite Zn-MoS with zinc doping2Gold-supported@platinum nanometer rods Au@Pt are substrate, then electropolymerization Shandong
ITO electro-conductive glass after minot is as electrochemical luminescence working electrode W1, reduced graphene carried titanium dioxide nano-film sol
rGO/TiO2The ITO electro-conductive glass modified jointly with stilbestrol antibody-solutions is as optical electro-chemistry working electrode W2, Ag/AgCl
Electrode is used as to electrode CE as reference electrode RE, platinum electrode, and four electrodes are inserted in same electrolytic cell jointly and form four
Electrode system, obtained four electrode system is placed in magazine, that is, the difunctional alkene based on Two-dimensional Composites structure is made
Female phenol is without enzyme sensor.
2. the preparation of difunctional stilbestrol without enzyme sensor according to claim 1 based on Two-dimensional Composites structure
Method, it is characterised in that the molybdenum disulfide composite Zn-MoS of described zinc doping2For the molybdenum disulfide doped with zinc ion
Two-dimension nano materials, described two-dimension nano materials are scraps of paper shape, and thickness is about 10 ~ 20 nm.
3. the preparation of difunctional stilbestrol without enzyme sensor according to claim 1 based on Two-dimensional Composites structure
Method, it is characterised in that described golden@platinum nanometer rods Au@Pt are the rod-like nano material of core shell structure, the core shell structure
Rod-like nano material is using gold nanorods as core, and nano platinum particle is the rod-like nano material of shell, and the gold nanorods are rods
Shape golden nanometer particle, length are 20 ~ 40 nm.
4. the preparation of difunctional stilbestrol without enzyme sensor according to claim 1 based on Two-dimensional Composites structure
Method, it is characterised in that described reduced graphene carried titanium dioxide nano-film sol rGO/TiO2Born for reduced graphene
The aqueous solution of carrying of titanium dioxide nanometer sheet, the titanium dioxide nanoplate be square sheet titanium dioxide nano-particle, the length of side
For 60 ~ 80 nm.
5. the preparation of difunctional stilbestrol without enzyme sensor according to claim 1 based on Two-dimensional Composites structure
Method, it is characterised in that by the relative insertion electrolytic cell of W1 and W2 conductive one side, W1 and W2 spacing are the cm of 0.5 cm ~ 1.5.
6. detection method of the stilbestrol without enzyme sensor prepared by preparation method according to claim 1, its feature exist
In described stilbestrol is applied to the detection of stilbestrol without enzyme sensor, and detecting step is：
（1）Stilbestrol solution to be measured is added dropwise on W2, the alkene on stilbestrol and W2 in stilbestrol solution to be measured
After female phenol antibody mediated immunity combines, remaining material is rinsed out, then the stilbestrol secondary antibody progress of hemin Hemin marks is added dropwise
It is immune to combine, the stilbestrol secondary antibody of unnecessary Hemin marks is rinsed out with reference to after, W2 to be measured is made；
（2）Using electrochemical workstation, in the electrolyte containing fixed concentration hydrogen peroxide, using step voltage on W1
Method applies step voltage to W1, and initial voltage is 0 v, and step voltage is 0.7 ~ 0.9 v, and snap time is 10 ~ 30 s；Together
When, when being used on W2 to be measured, m- current methods apply constant voltage to W2 to be measured, and voltage is 0 ~ 0.6 v；Electricity on W2 to be measured
Stream can increase with the concentration of stilbestrol in stilbestrol solution to be measured and accordingly increase, and be drawn according to gained electric current increment value
The concentration of stilbestrol in stilbestrol solution to be measured.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610148497.0A CN105758849B (en) | 2016-03-16 | 2016-03-16 | A kind of preparation method and application of the difunctional stilbestrol without enzyme sensor based on Two-dimensional Composites structure |
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