CN105784815B - A kind of preparation method and application of the hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source - Google Patents
A kind of preparation method and application of the hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source Download PDFInfo
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- CN105784815B CN105784815B CN201610148499.XA CN201610148499A CN105784815B CN 105784815 B CN105784815 B CN 105784815B CN 201610148499 A CN201610148499 A CN 201610148499A CN 105784815 B CN105784815 B CN 105784815B
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- 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
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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
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Abstract
The invention discloses a kind of preparation methods of the difunctional hydrogen peroxide sensor without peripheral hardware light source based on function nano material structure, prepared sensor operations are simple, easy to carry, detection is fast, at low cost, can be used for quick, the Sensitive Detection to hydrogen peroxide in the fields such as daily production and life.
Description
Technical field
The present invention relates to a kind of preparation method for detecting the sensor of hydrogen peroxide, the sensor integration electroluminescentization
It learns and shines and optical electro-chemistry dual function.Belong to Nano-function thin films and electrochemical biosensor analysis technical field.
Background technology
Hydrogen peroxide is a kind of oxidant, can be decomposed into water and oxygen under normal circumstances, but slow, is catalyzed when being added
Agent（Or biological enzyme）When, reaction speed is accelerated, it is possible thereby to be made instantly available the amount or catalyst of hydrogen peroxide（Or biological enzyme）'s
Amount, while hydrogen peroxide also often exists in the form of reaction intermediate in vivo, therefore, hydrogen peroxide is examined in medicine
The fields such as disconnected, clinical treatment, environment measuring and food production play important role, research and development hydrogen peroxide detection side
Method also just has highly important application value.Electrochemical biosensor analytical technology is fast etc. due to easy to operate, detection speed
Advantage increasingly obtains the attention of people.The electrochemical biosensor analytical technology of detection hydrogen peroxide is presently used for according to detection
Means have three kinds of electrochemical sensor, Electrochemiluminescsensor sensor and optical electro-chemistry sensor to divide mainly.Wherein, electroluminescentization
Luminescence sensor and optical electro-chemistry sensor are learned relative to electrochemical sensor, there is few background signal noise, high sensitivity, inspection
It surveys the features such as at low cost, it is of interest by more and more researchers in recent years.
Electrogenerated chemiluminescence is also referred to as electrochemical luminescence, refers to that generate some in electrode surface by electrochemical method special
Substance, form excitation state between these substances or by electron transmission between other components in system, returned by excitation state
Luminescence phenomenon is generated to ground state.Electrochemiluminescsensor sensor is the decorative material by changing electrode surface, with analysis produce
Raw electrochemical luminescence is realized according to the associated change of analyte concentration and electrochemical luminescence intensity to analysis in optimal conditions
The qualitative and quantitative analysis of object.
Optical electro-chemistry sensor is to cause electron-hole pair to be detached 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 variation of analyte concentration will have a direct impact on the size of photoelectric current, can be according to the change of photoelectric current
Change the qualitative and quantitative analysis realized to analyte.
But since Electrochemiluminescsensor sensor needs external optical signal to capture equipment such as photodiode etc., and light
Electrochemical sensor needs peripheral hardware light source to excite Electrophotosensitivmaterial material, this affects the convenient of the two operation to a certain extent
Property, it limits them and is more widely applied in actual production, life.Therefore, it designs, prepare more simple, fast inspection
The electrochemical biosensor analytical technology for surveying hydrogen peroxide has highly important practical value.
Invention content
The purpose of the present invention is to provide a kind of hydrogen peroxide sensings easy to operate, easy to carry, detection is fast, at low cost
The preparation method of device, prepared sensor, can be used for the fields such as daily production and life to the quick, sensitive of hydrogen peroxide
Detection.Based on this purpose, the present invention is in same electrolytic cell, and using four electrode systems, i.e. two working electrodes, one to electrode
With a reference electrode, wherein working electrode 1 is using gold vulcanization Silver nanorod Sol A u Ag2S NRs and electropolymerization luminol
It is modified jointly, as electrogenerated chemiluminescence working electrode W1, working electrode 2 is using titanium dioxide nanoplate colloidal sol TiO2
NSs and horseradish peroxidase Solution H RP are modified jointly, as optical electro-chemistry working electrode W2.When being detected, in electricity
After hydrogen peroxide is added in Xie Chizhong, apply step voltage on W1, due to Au Ag2The catalytic action of S NRs, luminol and mistake
Hydrogen oxide reacts, and generates electrogenerated chemiluminescence, this is just equivalent to " turning on light ", and when step voltage is 0, electrogenerated chemiluminescence disappears
It loses, this is just equivalent to " turning off the light ", at the same time applies constant voltage always on W2, due to TiO2NSs can be because of electroluminescent chemistry
The luminous excitation generated that shines causes electron-hole pair to be detached, and HRP catalyzing hydrogen peroxides generate oxygen, make hydrogen peroxide
As hole " donor ", to obtain photoelectric current on W2, when electrogenerated chemiluminescence disappears, i.e., when " turning off the light ", photoelectric current is immediately
It disappears, size and the concentration of hydrogen peroxide positive correlation of the photoelectric current of generation, therefore the size by recording photoelectric current can be realized
Detection to hydrogen peroxide.
Based on the above inventive principle, the specific technical solution that the present invention uses is as follows：
1. a kind of preparation method of the hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source, feature are, step is prepared
Suddenly it is：
（1）The preparation method of electrogenerated chemiluminescence working electrode W1, the W1 are to vulcanize Silver nanorod colloidal sol by gold
Au@Ag2The ITO electro-conductive glass that S NRs and electropolymerization luminol are modified jointly, its main feature is that, specific preparation process is：
1）Using ITO electro-conductive glass as working electrode, in electrode surface drop coating Au@Ag2S NRs, area coverage be 1 cm ×
1 cm, dries at room temperature；
2）By 1）Obtained working electrode immerses in electrolyte, and immersion area is Au@Ag2The area that S NRs are covered,
Electrochemical deposition is carried out to working electrode using three-electrode system, working electrode is taken out after deposition, is cleaned using ultra-pure water, 4 DEG C
Under be protected from light drying, electrogenerated chemiluminescence working electrode W1 is made；
The gold vulcanization Silver nanorod Sol A u Ag2S NRs are the aqueous solution of the rod-like nano material of nucleocapsid,
The rod-like nano material of the nucleocapsid is using gold nanorods as core, and vulcanization Nano silver grain is the rod-like nano material of shell
Material, the gold nanorods are bar-like gold nanoparticle, and length is 20 ~ 40 nm；
The electrolyte is the sulfuric acid solution containing luminol, a concentration of the 1 ~ 10 of luminol in the electrolyte
Mmol/L, sulfuric acid concentration are 0.1 ~ 1.0 mol/L；
The three-electrode system, including working electrode, reference electrode and to electrode, the reference electrode be saturated it is sweet
Mercury electrode, described is platinum electrode to electrode；
The electrochemical deposition process, the electrochemical method used is cyclic voltammetry, and starting voltage is -0.2 V, eventually
Only voltage is 1.5 V, and it is 100 mv/s, 20 ~ 30 circle of cycle to sweep speed；
（2）The preparation method of optical electro-chemistry working electrode W2, the W2 are by TiO2What NSs and HRP was modified jointly
ITO electro-conductive glass, its main feature is that, specific preparation process is：
1）Using ITO electro-conductive glass as working electrode, in 8 ~ 12 μ L TiO of electrode surface drop coating2NSs dries at room temperature；
2）By 1）In obtained working electrode be put into Muffle furnace, made annealing treatment at 450 DEG C, handle postcooling
To room temperature；
3）By 2）In obtained working electrode surface drop coating 8 ~ 12 μ L HRP, it is dry at 4 DEG C, it is clear with ultra-pure water after dry
It washes, it is dry at 4 DEG C, optical electro-chemistry working electrode W2 is made；
The TiO2NSs is the aqueous solution of the titanium dioxide nanoplate of 1 mg/mL, and the titanium dioxide nanoplate is
The titanium dioxide nano-particle of rectangular sheet, the length of side are 60 ~ 80 nm；
The HRP is the horseradish peroxidase aqueous solution of 5 mU/mL；
（3）The preparation method of hydrogen peroxide optical electro-chemistry sensor without peripheral hardware light source：
1）By in the opposite insertion electrolytic cell of the one side of W1 and W2 conductions, W1 is the cm of 0.5 cm ~ 1.5 with W2 spacing；
2）It is to be inserted into electrolytic cell to electrode CE by reference electrode RE, platinum electrode of Ag/AgCl, it is common with W1 and W2
Form four electrode systems；
3）The NaOH solution that 10 mL pH value are 11 ~ 13 is added in electrolytic cell；
4）By 1）~3）Obtained four electrode system and electrolytic cell are placed in magazine, obtain the peroxide of no peripheral hardware light source
Change hydrogen optical electro-chemistry sensor.
2. the detection applied to hydrogen peroxide of hydrogen peroxide optical electro-chemistry sensor of the present invention, its main feature is that
Specifically detection method is：
（1）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 when being used on W2 m- current methods to W2
Apply constant voltage, voltage is 0 ~ 0.6 v；
（2）The Hydrogen peroxide standard solution of various concentration is added in electrolytic cell, the electric current on W2 can be with hydrogen peroxide
The increase of concentration and accordingly increase, according to the relationship between gained electric current increment value and concentration of hydrogen peroxide, drawing curve；
（3）The standard solution that hydrogenperoxide steam generator to be measured is replaced to hydrogen peroxide, according to（1）With（2）The peroxidating
Hydrogen detection method is detected, and show that hydrogen peroxide to be measured is molten with the working curve drawn according to obtained electric current increment value
The concentration of liquid.
The useful achievement of the present invention
（1）Hydrogen peroxide sensor of the present invention is prepared simply, easy to operate, is not necessarily to external accessory, is utilized
The micromation of detection device, portability, and quick, sensitive, the highly selective detection to hydrogen peroxide is realized, have wide
Market development foreground；
（2）The present invention detects hydrogen peroxide in same electrolytic cell using four electrode systems for the first time, and realizes electroluminescentization
It learns and shines and the difunctional signal amplification strategy of optical electro-chemistry.With the increase of concentration of hydrogen peroxide in electrolytic cell, on the one hand, make
Obtain the photoelectricity cleanliness increase that electrogenerated chemiluminescence intensity is linearly increasing, is excited；On the other hand, hydrogen peroxide is given as electronics
Body so that photoelectricity cleanliness increases in optical electro-chemistry reaction；The third aspect, due to Au@Ag2The big specific surface area of S NRs with
And and luminol amino combination, electropolymerization luminol can be made preferably to load to electrode surface, increase luminous intensity.Cause
This, may be implemented two methods of electrogenerated chemiluminescence and optical electro-chemistry and be total in same electrolytic cell, under same electrochemical workstation
With reaction, interaction, realize to hydrogen peroxide detect electric signal dual amplification, greatly improve detection sensitivity and
Detection limit has important scientific meaning.Since the photoelectric sensor is not necessarily to peripheral hardware light source, it is more advantageous to micromation, portability,
So having a vast market application value.
Specific implementation mode
A kind of hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source of embodiment 1, specific preparation process are：
（1）The preparation of electrogenerated chemiluminescence working electrode W1：
1）Using ITO electro-conductive glass as working electrode, in electrode surface drop coating Au@Ag2S NRs, area coverage be 1 cm ×
1 cm, dries at room temperature；
2）By 1）Obtained working electrode immerses in electrolyte, and immersion area is Au@Ag2The area that S NRs are covered,
Electrochemical deposition is carried out to working electrode using three-electrode system, working electrode is taken out after deposition, is cleaned using ultra-pure water, 4 DEG C
Under be protected from light drying, electrogenerated chemiluminescence working electrode W1 is made；
The Au@Ag2S NRs be nucleocapsid rod-like nano material aqueous solution, the nucleocapsid it is rodlike
Nano material is using gold nanorods as core, and vulcanization Nano silver grain is the rod-like nano material of shell, and the gold nanorods are sticks
Shape gold nanoparticle, length are 20 nm；
The electrolyte is the sulfuric acid solution containing luminol, a concentration of the 1 of luminol in the electrolyte
Mmol/L, sulfuric acid concentration are 0.1 mol/L；
The three-electrode system, including working electrode, reference electrode and to electrode, the reference electrode be saturated it is sweet
Mercury electrode, described is platinum electrode to electrode；
The electrochemical deposition process, the electrochemical method used is cyclic voltammetry, and starting voltage is -0.2V, eventually
Only voltage is 1.5V, and it is 100mv/s, 20 circle of cycle to sweep speed.
（2）The preparation of optical electro-chemistry working electrode W2：
1）Using ITO electro-conductive glass as working electrode, in 8 μ L TiO of electrode surface drop coating2NSs dries at room temperature；
2）By 1）In obtained working electrode be put into Muffle furnace, made annealing treatment at 450 DEG C, handle postcooling
To room temperature；
3）By 2）In obtained working electrode surface drop coating 8 μ L HRP, it is dry at 4 DEG C, cleaned with ultra-pure water after dry,
It is dry at 4 DEG C, optical electro-chemistry working electrode W2 is made；
The TiO2NSs is the aqueous solution of the titanium dioxide nanoplate of 1 mg/mL, and the titanium dioxide nanoplate is
The titanium dioxide nano-particle of rectangular sheet, the length of side are 60 ~ 80 nm；
The HRP is the horseradish peroxidase aqueous solution of 5 mU/mL.
（3）The preparation method of hydrogen peroxide optical electro-chemistry sensor without peripheral hardware light source：
1）It will（1）The W1 of middle preparation and（2）The W2 of middle preparation is inserted into electrolytic cell face to face, and W1 is 0.5 with W2 spacing
cm；
2）It is to be inserted into electrolytic cell to electrode CE by reference electrode RE, platinum electrode of Ag/AgCl, it is common with W1 and W2
Form four electrode systems；
3）The NaOH solution that 10 mL pH value are 11 is added in electrolytic cell；
4）By 1）~3）Obtained four electrode system and electrolytic cell are placed in magazine, obtain the peroxide of no peripheral hardware light source
Change hydrogen optical electro-chemistry sensor.
A kind of hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source of embodiment 2, specific preparation process are：
（1）The preparation of electrogenerated chemiluminescence working electrode W1：
For preparation process with the preparation process of W1 in embodiment 1, difference is that the length of gold nanorods is 30 nm, electrolysis
A concentration of 5 mmol/L of luminol in liquid, sulfuric acid concentration are that 0.5 mol/L is followed when cyclic voltammetry carries out electrochemical deposition
Ring 25 encloses.
（2）The preparation of optical electro-chemistry working electrode W2：
1）Using ITO electro-conductive glass as working electrode, in 10 μ L TiO of electrode surface drop coating2NSs dries at room temperature；
2）By 1）In obtained working electrode be put into Muffle furnace, made annealing treatment at 450 DEG C, handle postcooling
To room temperature；
3）By 2）In obtained working electrode surface drop coating 10 μ L HRP, it is dry at 4 DEG C, it is clear with ultra-pure water after dry
It washes, it is dry at 4 DEG C, optical electro-chemistry working electrode W2 is made；
Remaining preparation process with W2 in embodiment 1.
（3）The preparation method of hydrogen peroxide optical electro-chemistry sensor without peripheral hardware light source：
Preparation process is with embodiment 1, and difference is W1 and W2 spacing is 1.0 cm, and the NaOH being added in electrolytic cell is molten
The pH value of liquid is 12.
A kind of hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source of embodiment 3, specific preparation process are：
（1）The preparation of electrogenerated chemiluminescence working electrode W1：
For preparation process with the preparation process of W1 in embodiment 1, difference is that the length of gold nanorods is 40 nm, electrolysis
A concentration of 10 mmol/L of luminol in liquid, sulfuric acid concentration are that 1.0 mol/L are followed when cyclic voltammetry carries out electrochemical deposition
Ring 30 encloses.
（2）The preparation of optical electro-chemistry working electrode W2：
1）Using ITO electro-conductive glass as working electrode, in 12 μ L TiO of electrode surface drop coating2NSs dries at room temperature；
2）By 1）In obtained working electrode be put into Muffle furnace, made annealing treatment at 450 DEG C, handle postcooling
To room temperature；
3）By 2）In obtained working electrode surface drop coating 12 μ L HRP, it is dry at 4 DEG C, it is clear with ultra-pure water after dry
It washes, it is dry at 4 DEG C, optical electro-chemistry working electrode W2 is made；
Remaining preparation process with W2 in embodiment 1.
（3）The preparation method of hydrogen peroxide optical electro-chemistry sensor without peripheral hardware light source：
Preparation process is with embodiment 1, and difference is W1 and W2 spacing is 1.5 cm, and the NaOH being added in electrolytic cell is molten
The pH value of liquid is 13.
A kind of application of the hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source of embodiment 4
A kind of hydrogen peroxide optical electro-chemistry sensor application of no peripheral hardware light source prepared by embodiment 1 is in the inspection of hydrogen peroxide
It surveys, detecting step is：
（1）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 m- current methods are constant to W2 applications when being used on W2
Voltage, voltage are 0 v；
（2）The Hydrogen peroxide standard solution of various concentration is added in electrolytic cell, the electric current on W2 can be with hydrogen peroxide
The increase of concentration and accordingly increase, according to the relationship between gained electric current increment value and concentration of hydrogen peroxide, drawing curve；
（3）The standard solution that hydrogenperoxide steam generator to be measured is replaced to hydrogen peroxide, according to（1）With（2）The peroxidating
Hydrogen detection method is detected, and show that hydrogen peroxide to be measured is molten with the working curve drawn according to obtained electric current increment value
The concentration of liquid.
A kind of application of the hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source of embodiment 5
A kind of hydrogen peroxide optical electro-chemistry sensor application of no peripheral hardware light source prepared by embodiment 2 is in the inspection of hydrogen peroxide
It surveys, detecting step is：
（1）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 0.8 v, and snap time is 20 s；Meanwhile m- current methods are constant to W2 applications when being used on W2
Voltage, voltage are 0.3 v；
（2）With（3）With embodiment 4.
A kind of application of the hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source of embodiment 6
A kind of hydrogen peroxide optical electro-chemistry sensor application of no peripheral hardware light source prepared by embodiment 3 is in the inspection of hydrogen peroxide
It surveys, detecting step is：
（1）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 0.9 v, and snap time is 30 s；Meanwhile m- current methods are constant to W2 applications when being used on W2
Voltage, voltage are 0.6 v；
（2）With（3）With embodiment 4.
Hydrogen peroxide optical electro-chemistry sensor prepared by 7 embodiment 1-3 of embodiment is applied to the inspection of hydrogen peroxide
It surveys, there is excellent detection result, detection to be limited to 12 μm of ol/L.
The detection of hydrogen peroxide in 8 human serum of embodiment
Human serum sample accurately is pipetted, the Hydrogen peroxide standard solution of certain molar concentration is added, peroxidating not to be added
The human serum of hydrogen is blank, carries out recovery testu, is detected according to the step of embodiment 4 ~ 6, peroxide in determination sample
Change the rate of recovery of hydrogen, testing result is shown in Table 1.
The testing result of hydrogen peroxide in 1 human serum of table
1 testing result of table it is found that result relative standard deviation（RSD）Less than 2.8 %, the rate of recovery is 94 ~ 106%, table
The bright present invention can be used for the detection of hydrogen peroxide in human serum, the high sensitivity of method, high specificity, as a result accurately and reliably.
Claims (5)
1. a kind of preparation method of the hydrogen peroxide optical electro-chemistry sensor of no peripheral hardware light source, which is characterized in that use gold@sulphur
Change Silver nanorod Sol A u@Ag2The ITO electro-conductive glass that S NRs and electropolymerization luminol are modified jointly is as electrogenerated chemiluminescence
Working electrode W1, titanium dioxide nanoplate colloidal sol TiO2The ITO that NSs and horseradish peroxidase Solution H RP are modified jointly is conductive
Glass is used as reference electrode RE, platinum electrode to electrode CE as optical electro-chemistry working electrode W2, Ag/AgCl electrode, will
Four electrodes, which are inserted into jointly in same electrolytic cell, forms four electrode systems, and four electrode system obtained is placed in magazine, that is, is made
Obtain the hydrogen peroxide optical electro-chemistry sensor without peripheral hardware light source.
2. preparation method according to claim 1, which is characterized in that the gold vulcanization Silver nanorod Sol A u Ag2S
NRs is the aqueous solution of the rod-like nano material of nucleocapsid, and the rod-like nano material of the nucleocapsid is to be with gold nanorods
Core, vulcanization Nano silver grain are the rod-like nano material of shell, and the gold nanorods are bar-like gold nanoparticle, length is 20 ~
40 nm。
3. preparation method according to claim 1, which is characterized in that the titanium dioxide nanoplate colloidal sol TiO2 NSs
For the aqueous solution of titanium dioxide nanoplate, the titanium dioxide nanoplate is the titanium dioxide nano-particle of rectangular sheet, the length of side
For 60 ~ 80 nm.
4. preparation method according to claim 1, which is characterized in that the one side of W1 and W2 conductions is inserted into electrolysis relatively
Chi Zhong, W1 and W2 spacing are the cm of 0.5 cm ~ 1.5.
5. hydrogen peroxide optical electro-chemistry sensor prepared by preparation method according to claim 1, which is characterized in that institute
The hydrogen peroxide optical electro-chemistry sensor application stated is the step of hydrogen peroxide detects：Using electrochemical workstation, on W1
Step voltage is applied to W1 using the method for step voltage, initial voltage is 0 v, and step voltage is 0.7 ~ 0.9 v, snap time
For 10 ~ 30 s；Meanwhile m- current methods apply constant voltage to W2 when being used on W2, voltage is 0 ~ 0.6 v；In electrolytic cell
Middle that hydrogenperoxide steam generator to be measured is added, the electric current on W2 can accordingly increase with the increase of concentration of hydrogen peroxide, according to gained
Electric current increment value obtains hydrogenperoxide steam generator concentration to be measured.
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| CN203824914U (en) * | 2014-04-17 | 2014-09-10 | 苏州大学 | Double-constant -potential excited electrochemical luminescence analysis device |
| CN104155445A (en) * | 2014-07-14 | 2014-11-19 | 济南大学 | Preparation and application of unmarked electrochemiluminescent tumor marker immunosensor |
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| CN203824914U (en) * | 2014-04-17 | 2014-09-10 | 苏州大学 | Double-constant -potential excited electrochemical luminescence analysis device |
| CN104155445A (en) * | 2014-07-14 | 2014-11-19 | 济南大学 | Preparation and application of unmarked electrochemiluminescent tumor marker immunosensor |
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