CN106053566B - A kind of preparation method of the optical electro-chemistry salbutamol sensor based on two-dimensional magnetic nanometer photoelectronic material - Google Patents
A kind of preparation method of the optical electro-chemistry salbutamol sensor based on two-dimensional magnetic nanometer photoelectronic material Download PDFInfo
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Abstract
The invention discloses a kind of preparation methods of optical electro-chemistry salbutamol sensor.Belong to Nano-function thin films and biosensor technology field.The method comprises the steps of firstly, preparing a kind of New Two Dimensional magnetic Nano photoelectric materials, i.e. the two-dimensional nano composite material Fe-TiO of Fe2O3 doping nano titania square In-situ reaction molybdenum disulfide2/MoS2Good biocompatibility and big specific surface area using the material, salbutamol antibody in load, alkaline phosphatase in fixation, when being detected, L-AA AA is generated in situ since alkaline phosphatase can be catalyzed L-AA -2- tricresyl phosphate sodium salt AAP, and electron donor is provided for Photoelectric Detection in turn, recycle antibody in conjunction with the specific quantification of antigen to the influence of electron transport ability, so that photo-current intensity accordingly reduces, the building of the photoelectric sensor using unmarked PhotoelectrochemicalMethod Method detection salbutamol is finally realized.
Description
Technical field
The present invention relates to a kind of preparation methods of optical electro-chemistry salbutamol sensor.Belong to Nano-function thin films with
Biosensor technology field.
Background technique
Salbutamol is selective β 2 receptor agonist, is used as clenbuterol hydrochloride, Lai Tigao live pig in the cultivation of modern pork
Lean meat yield.Begin within 2002, is listed in cultivation industry forbidden drug, must not add in livestock and poultry breeding.
Currently, the method for detection salbutamol mainly has chromatography, mass spectrography etc..Such method instrument is valuable, operation is multiple
Miscellaneous, laboratory personnel just can be carried out detection after needing professional training.Therefore, research and development are at low cost, detection is fast, high sensitivity, specificity
Strong salbutamol sensor is of great significance.
Optical electro-chemistry sensor due to high sensitivity, testing cost is low the features such as, in recent years by more and more researchers
It is of interest.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 transmitting of the electronics on electrode, semiconductor and modifier and analyte is realized, and form light
Electric current.In optimal conditions, the variation of analyte concentration will have a direct impact on the size of photoelectric current, and biological immune is recycled to combine,
The qualitative and quantitative analysis to analyte can be realized according to the variation of photoelectric current.
Optical electro-chemistry sensor most critical technology is exactly the raising to performances such as the size of photoelectric current and stability.Titanium dioxide
Titanium is a kind of photochemical catalyst and light induced electron host material being most widely used, however, to give full play to the reality of titanium dioxide
Border application level needs on the one hand living to improve optical electro-chemistry with the more high activity crystal faces of exposure by regulating and controlling its material morphology
Property, photosensitive wavelength is on the other hand regulated and controled by doping different metal or metal oxide and improves the sun to visible-range extension
The utilization rate of light.It, can due to two-dimentional titanium dioxide nano material, such as titanium dioxide nanoplate, nano titania square
The more high activity crystal faces of exposure have higher optical electro-chemistry activity, and titanium dioxide nanoplate has more preferable than nanoparticle
Ground application prospect is also concerned the research of titanium dioxide nanoplate.And single titanium dioxide nano material is photosensitive
Wavelength generally in ultra-violet (UV) band, and due to bad dispersibility, easily stack and interact, thus reduce optical electro-chemistry activity, it is unfavorable
In practical application.Therefore, at low cost, preparing simple bloom electro-chemical activity photosensitive dose of titanium dioxide is researched and developed with important
Scientific meaning and application value.
Molybdenum disulfide (chemical formula MoS2) nano material, there is two-dimensional layered structure, be most widely used solid profit
One of lubrication prescription.Its sheet two-dimension nano materials after removing, is the semiconductor nano material haveing excellent performance, in addition to big ratio
Surface area, can be used as the carrier of catalyst and biological antibody, improve load capacity, while also have as co-catalyst excellent
Electron transmission performance.
Currently, most of synthesizing mean is all after being separately synthesized, then catalyst and carrier progress is compound, process is numerous
Trivial, yield is not high.Therefore, before being had a wide range of applications for In-situ reaction preparation with the photosensitizer of excellent photoelectrochemical behaviour
Scape and important scientific meaning.
In addition, the photo-generate electron-hole of single titanium dioxide nano material to easily it is compound, so as to cause photosignal
Weaken, and titanium dioxide poorly conductive also limits the optical electro-chemistry sensor constructed by single titanium dioxide nano material
Sensitivity is not generally high, is unfavorable for practical application.Therefore, design, prepare efficient, stable doping titanium dioxide nano piece and its
Modifier is the key technology for preparing optical electro-chemistry sensor.
Summary of the invention
The purpose of the present invention is to provide it is a kind of prepare simple, high sensitivity, detection quickly, the optical electro-chemistry of high specificity
The preparation method of salbutamol sensor, prepared sensor can be used for quick, the Sensitive Detection of salbutamol.Based on this
Purpose, the method comprises the steps of firstly, preparing a kind of New Two Dimensional magnetic Nano photoelectric material, i.e. Fe2O3 doping nano titania square is former
The two-dimensional nano composite material Fe-TiO of position composite molybdenum disulfide2/MoS2, utilize the good biocompatibility of the material and big
Specific surface area, salbutamol antibody in load, alkaline phosphatase in fixation, when being detected, since alkaline phosphatase can
L-AA AA is generated in situ to be catalyzed L-AA -2- tricresyl phosphate sodium salt AAP, and provides electronics in turn for Photoelectric Detection
Donor recycles antibody in conjunction with the specific quantification of antigen to the influence of electron transport ability, so that photo-current intensity is corresponding
It reduces, finally realizes the building of the photoelectric sensor using unmarked PhotoelectrochemicalMethod Method detection salbutamol.
The technical solution adopted by the invention is as follows:
1. a kind of preparation method of the optical electro-chemistry salbutamol sensor based on two-dimensional magnetic nanometer photoelectronic material,
It is characterized in that the two-dimensional magnetic nanometer photoelectronic material is Fe2O3 doping nano titania square In-situ reaction molybdenum disulfide
Two-dimensional nano composite material Fe-TiO2/MoS2, the optical electro-chemistry salbutamol sensor is by working electrode, Fe-TiO2/
MoS2, salbutamol antibody, alkaline phosphatase, bovine serum albumin(BSA) composition；
It is characterized in that, the preparation method includes following preparation step:
A. Fe-TiO is prepared2/MoS2；
B. optical electro-chemistry salbutamol sensor is prepared；
Wherein, step a prepares Fe-TiO2/MoS2Specific steps are as follows:
(1) 0.6 g molybdenum disulfide powder and 0.2 ~ 2.0 mmol molysite is taken to be added to 3 ~ 10 mL n-BuLis jointly molten
In liquid, at nitrogen protection and 30 ~ 60 DEG C, stir 12 ~ 48 hours, the solution after being reacted；
(2) using the solution after reaction in nonpolar solvent washing step (1), water-bath is then carried out at 30 ~ 60 DEG C
Ultrasonic treatment, after handle, recycle nonpolar solvent carrying out washing treatment after solution, be dried in vacuo, obtain two sulphur of iron intercalation
Change molybdenum nano material；
(3) the molybdenum disulfide nano material of iron intercalation made from 10 ~ 500 mg steps (2) is taken to be added to 5 mL metatitanic acids four
In butyl ester, after stirring 1 hour, it is slowly added to 0.5 ~ 0.8 mL hydrofluoric acid while stirring, is then reacting at 160 ~ 180 DEG C
It is reacted 18 ~ 20 hours in kettle；
(4) by step (3) resulting reaction product, with ultrapure water and dehydrated alcohol centrifuge washing three times after, it is true at 50 DEG C
Sky is dry, obtains Fe-TiO2/MoS2；
The n-butyllithium solution is the hexane solution of n-BuLi, and concentration is 1.6 mol/L；
The molysite is selected from one of following: ferric sulfate, iron chloride, ferric nitrate, organoiron compound；
The nonpolar solvent is selected from one of following: hexane, hexamethylene, carbon tetrachloride, benzene, toluene；
The water bath sonicator processing, processing time are 1 hour；
The specific steps of step b preparation optical electro-chemistry salbutamol sensor are as follows:
(1) using ITO electro-conductive glass as working electrode, in the Fe-TiO of 8 ~ 12 μ L of electrode surface drop coating2/MoS2Colloidal sol, room
It is dried under temperature；
(2) electrode obtained in step (1) is cleaned with buffer solution PBS, is continued in 8 ~ 12 μ L of electrode surface drop coating
The salbutamol antibody-solutions of 10 μ g/mL are saved in 4 DEG C of refrigerators and are dried；
(3) electrode obtained in step (2) is cleaned with PBS, continues in 6 ~ 10 μ L concentration of electrode surface drop coating to be 20
The alkaline phosphatase enzyme solutions of μ g/mL are saved in 4 DEG C of refrigerators and are dried；
(4) electrode obtained in step (3) is cleaned with PBS, continues in 8 ~ 12 μ L concentration of electrode surface drop coating to be 100
The bovine serum albumin solution of μ g/mL is saved in 4 DEG C of refrigerators and is dried；
(5) electrode obtained in step (4) is cleaned with PBS, is saved in 4 DEG C of refrigerators after drying, obtains photoelectricity
Chemical salbutamol sensor；
The Fe-TiO2/MoS2Colloidal sol is by the Fe-TiO of 50 mg2/MoS2Powder is dissolved in 10 mL ultrapure waters, and
The hydrosol obtained after 30 min of ultrasound；
The PBS is the phosphate buffer solution of 10 mmol/L, and the pH value of the phosphate buffer solution is 7.4.
2. the application of optical electro-chemistry salbutamol sensor, feature prepared by preparation method of the present invention exist
In, including following applying step:
A. standard solution is prepared: preparing the salbutamol standard solution of one group of various concentration including blank standard specimen；
B. working electrode is modified: optical electro-chemistry salbutamol prepared by preparation method as described in claim 1 is passed
Sensor is working electrode, and the salbutamol standard solution for the various concentration prepared in step b is distinguished drop coating to working electrode table
Face saves in 4 DEG C of refrigerators；
C. working curve is drawn: using saturated calomel electrode as reference electrode, platinum electrode is as auxiliary electrode, with step
The working electrode composition three-electrode system that rapid b has been modified, is connected on optical electro-chemistry detection device；Successively add in a cell
L-AA -2- tricresyl phosphate sodium salt the AAP for entering 10 mmol/L of Tris-HCl buffer solution and 5 mL of pH=9.6 15mL is molten
Liquid；Work is drawn according to the relationship between resulting photocurrent values and salbutamol concentration of standard solution using i-t means of testing
Make curve；
D. the detection of salbutamol: replacing the salbutamol standard solution in step a with sample to be tested, according to step b and
Method in c is detected, according to response the intensity value and working curve of signal, obtains containing for salbutamol in sample to be tested
Amount.
Beneficial achievement of the invention
(1) optical electro-chemistry salbutamol sensor preparation of the present invention is simple, easy to operate, realizes to sample
Quickly, sensitive, highly selective detection, and it is at low cost, it can be applied to portable inspectiont, there is market development prospect；
(2) present invention is prepared for novel photocatalyst Fe-TiO using the method for In-situ reaction for the first time2/MoS2, this method master
Will be there are three advantage: first is that, due to growth in situ of the iron on nano titania square sufficiently with nano titania side
Block contact is acted on using the metal surface plasma body of iron, effectively prevents the compound of photo-generate electron-hole pair, greatly mention
High photocatalytic activity, due to the effect of metal ion, range, realizes in visible region light and urges with having widened photosensitive wavelength
Change acts on, with greatly improving sunlight utilization efficiency, although solving two-dimentional titanium dioxide nano material photocatalysis effect
It is good, but under sunlight irradiation photocatalysis effect difference technical problem；Second is that due to molybdenum disulfide sheet two-dimension nano materials
Load characteristic and nano titania square on it fully dispersed, greatly increase the light of nano titania square
It catalytic activity and solves two-dimentional titanium dioxide nano material and is unfavorable for dispersing and the technical issues of reduce photocatalytic activity；Three
It is, since iron ion is in this process not only as intercalation material but also as reaction dopant material, finally to use the side of In-situ reaction
Method realizes one pot of preparation of the composite material, not only saves time, material loss, and make the two of the Fe2O3 doping prepared
TiOx nano square can preferably be evenly spread to above molybdenum disulfide sheet two-dimension nano materials.Therefore, the material
Effectively preparation has important scientific meaning and application value；
(3) present invention is for the first time by Fe-TiO2/MoS2Applied in the preparation of Photoelectrochemistrbiosensor biosensor, significantly improve
The effective concentration of photo-generated carrier substantially increases the detection sensitivity of optical electro-chemistry sensor, so that optical electro-chemistry biology passes
Sensor realizes application in actual operation；The application of the material is also associated biomolecule sensor, as electrogenerated chemiluminescence passes
Sensor, electrochemical sensor etc. provide Technical Reference, have extensive potential use value.
Specific embodiment
1 Fe-TiO of embodiment2/MoS2Preparation
(1) 0.6 g molybdenum disulfide powder and 0.2 mmol molysite is taken to be added in 3mL n-butyllithium solution jointly, in nitrogen
At gas shielded and 60 DEG C, stir 12 hours, the solution after being reacted；
(2) using the solution after reaction in nonpolar solvent washing step (1), water bath sonicator is then carried out at 60 DEG C
Processing, after handle, recycle nonpolar solvent carrying out washing treatment after solution, be dried in vacuo, obtain the molybdenum disulfide of iron intercalation
Nano material；
(3) the molybdenum disulfide nano material of iron intercalation made from 500 mg steps (2) is taken to be added to 5 mL butyl titanates
In, after stirring 1 hour, it is slowly added to 0.5 mL hydrofluoric acid while stirring, then reacts in a kettle 18 hours at 160 DEG C；
(4) by step (3) resulting reaction product, with ultrapure water and dehydrated alcohol centrifuge washing three times after, it is true at 50 DEG C
Sky is dry, obtains Fe-TiO2/MoS2；
The n-butyllithium solution is the hexane solution of n-BuLi, and concentration is 1.6 mol/L；
The molysite is ferric sulfate；
The nonpolar solvent is hexane；
The water bath sonicator processing, processing time are 1 hour.
2 Fe-TiO of embodiment2/MoS2Preparation
(1) 0.6 g molybdenum disulfide powder and 1.0 mmol molysite is taken to be added in 5 mL n-butyllithium solutions jointly, in nitrogen
At gas shielded and 30 DEG C, stir 24 hours, the solution after being reacted；
(2) using the solution after reaction in nonpolar solvent washing step (1), water bath sonicator is then carried out at 30 DEG C
Processing, after handle, recycle nonpolar solvent carrying out washing treatment after solution, be dried in vacuo, obtain the molybdenum disulfide of iron intercalation
Nano material；
(3) the molybdenum disulfide nano material of iron intercalation made from 200 mg steps (2) is taken to be added to 5 mL butyl titanates
In, after stirring 1 hour, it is slowly added to 0.6 mL hydrofluoric acid while stirring, then reacts in a kettle 20 hours at 180 DEG C；
(4) by step (3) resulting reaction product, with ultrapure water and dehydrated alcohol centrifuge washing three times after, it is true at 50 DEG C
Sky is dry, obtains Fe-TiO2/MoS2；
The n-butyllithium solution is the hexane solution of n-BuLi, and concentration is 1.6 mol/L；
The molysite is iron chloride；
The nonpolar solvent is carbon tetrachloride；
The water bath sonicator processing, processing time are 1 hour.
The preparation method of 3 optical electro-chemistry salbutamol sensor of embodiment
(1) using the ITO electro-conductive glass that width is 1 cm, a length of 4 cm as working electrode, 8 μ L's of electrode surface drop coating
Fe-TiO2/MoS2Colloidal sol dries at room temperature；
(2) electrode obtained in step (1) is cleaned with buffer solution PBS, is continued in 8 μ L of electrode surface drop coating, 10 μ
The salbutamol antibody-solutions of g/mL are saved in 4 DEG C of refrigerators and are dried；
(3) electrode obtained in step (2) is cleaned with PBS, continues in 8 μ L concentration of electrode surface drop coating to be 100 μ
The bovine serum albumin solution of g/mL is saved in 4 DEG C of refrigerators and is dried；
(4) electrode obtained in step (3) is cleaned with PBS, continues in 6 μ L concentration of electrode surface drop coating to be 20 μ g/
The alkaline phosphatase enzyme solutions of mL are saved in 4 DEG C of refrigerators and are dried；
(5) electrode obtained in step (4) is cleaned with PBS, is saved in 4 DEG C of refrigerators after drying, obtains photoelectricity
Chemical salbutamol sensor；
The Fe-TiO2/MoS2Colloidal sol is Fe-TiO prepared by the embodiment 1 by 50 mg2/MoS2Powder is dissolved in
In 10 mL ultrapure waters, and the hydrosol obtained after 30 min of ultrasound；
The PBS is the phosphate buffer solution of 10mmol/L, and the pH value of the phosphate buffer solution is 7.4.
The preparation method of 4 optical electro-chemistry salbutamol sensor of embodiment
All preparation steps are the same as embodiment 4, only Fe-TiO used in step2/MoS2For Fe- prepared by embodiment 2
TiO2/MoS2。
The optical electro-chemistry salbutamol sensor of 5 embodiment 1 and 3 of embodiment preparation, applied to the detection of salbutamol,
Steps are as follows:
(1) standard solution is prepared: preparing the salbutamol standard solution of one group of various concentration including blank standard specimen；
(2) working electrode is modified: optical electro-chemistry salbutamol prepared by preparation method as described in claim 1 is passed
Sensor is working electrode, and the salbutamol standard solution for the various concentration prepared in step (1) is distinguished drop coating to working electrode
Surface saves in 4 DEG C of refrigerators；
(3) working curve is drawn: using saturated calomel electrode as reference electrode, platinum electrode is as auxiliary electrode, with step
Suddenly the working electrode that (2) have been modified forms three-electrode system, is connected on optical electro-chemistry detection device；In a cell successively
L-AA -2- tricresyl phosphate sodium salt the AAP of 10 mmol/L of Tris-HCl buffer solution and 5 mL of pH=9.6 15mL is added
Solution；It is drawn using i-t means of testing according to the relationship between resulting photocurrent values and salbutamol concentration of standard solution
Working curve；The linear detection range of salbutamol are as follows: 0.002 ~ 200 ng/mL, detection limit are as follows: 0.8 pg/mL；
(4) actual sample detects: the salbutamol standard solution in step (1) is replaced with sample to be tested, according to step (2)
(3) method in is detected, and the intensity value and working curve of signal, obtain salbutamol in sample to be tested according to response
Content.
The optical electro-chemistry salbutamol sensor of 6 embodiment 2 and 4 of embodiment preparation, applied to the detection of salbutamol,
Steps are as follows:
(1) standard solution is prepared: preparing the salbutamol standard solution of one group of various concentration including blank standard specimen；
(2) working electrode is modified: optical electro-chemistry salbutamol prepared by preparation method as described in claim 1 is passed
Sensor is working electrode, and the salbutamol standard solution for the various concentration prepared in step (1) is distinguished drop coating to working electrode
Surface saves in 4 DEG C of refrigerators；
(3) working curve is drawn: using saturated calomel electrode as reference electrode, platinum electrode is as auxiliary electrode, with step
Suddenly the working electrode that (2) have been modified forms three-electrode system, is connected on optical electro-chemistry detection device；In a cell successively
L-AA -2- tricresyl phosphate sodium salt the AAP of 10 mmol/L of Tris-HCl buffer solution and 5 mL of pH=9.6 15mL is added
Solution；It is drawn using i-t means of testing according to the relationship between resulting photocurrent values and salbutamol concentration of standard solution
Working curve；The linear detection range of salbutamol are as follows: 0.002 ~ 200 ng/mL, detection limit are as follows: 0.8 pg/mL；
(4) actual sample detects: the salbutamol standard solution in step (1) is replaced with sample to be tested, according to step (2)
(3) method in is detected, and the intensity value and working curve of signal, obtain salbutamol in sample to be tested according to response
Content.
Claims (1)
1. a kind of preparation method of the optical electro-chemistry salbutamol sensor based on two-dimensional magnetic nanometer photoelectronic material, feature exist
In two wieners that the two-dimensional magnetic nanometer photoelectronic material is Fe2O3 doping nano titania square In-situ reaction molybdenum disulfide
Nano composite material Fe-TiO2/MoS2, the optical electro-chemistry salbutamol sensor is by working electrode, Fe-TiO2/MoS2, it is husky
Butylamine alcohol antibody, alkaline phosphatase, bovine serum albumin(BSA) composition；
The Fe-TiO2/MoS2Preparation step are as follows: take 0.6 g molybdenum disulfide powder and 0.2 ~ 2.0 mmol molysite common
It is added in 3 ~ 10 mL n-butyllithium solutions, at nitrogen protection and 30 ~ 60 DEG C, stirs 12 ~ 48 hours, reacted
Solution afterwards；Using the solution after nonpolar solvent washing reaction, water bath sonicator processing, place are then carried out at 30 ~ 60 DEG C
After having managed, recycle nonpolar solvent carrying out washing treatment after solution, vacuum drying, obtain the molybdenum disulfide nano material of iron intercalation
Material；The molybdenum disulfide nano material of iron intercalation made from 10 ~ 500 mg is taken to be added in 5 mL butyl titanates, stirring 1 is small
Shi Hou is slowly added to 0.5 ~ 0.8 mL hydrofluoric acid while stirring, then reacts 18 ~ 20 in a kettle at 160 ~ 180 DEG C
Hour, resulting reaction product, with ultrapure water and dehydrated alcohol centrifuge washing three times after, be dried in vacuo, obtain at 50 DEG C
Fe-TiO2/MoS2；
The n-butyllithium solution is the hexane solution of n-BuLi, and concentration is 1.6 mol/L；
The molysite is selected from one of following: ferric sulfate, iron chloride, ferric nitrate, organoiron compound；
The nonpolar solvent is selected from one of following: hexane, hexamethylene, carbon tetrachloride, benzene, toluene；
The water bath sonicator processing, processing time are 1 hour；
The preparation step of the optical electro-chemistry salbutamol sensor are as follows:
(1) using ITO electro-conductive glass as working electrode, in the Fe-TiO of 8 ~ 12 μ L of electrode surface drop coating2/MoS2Colloidal sol, at room temperature
It dries；
(2) electrode obtained in step (1) is cleaned with buffer solution PBS, is continued in 8 ~ 12 μ L of electrode surface drop coating, 10 μ
The salbutamol antibody-solutions of g/mL are saved in 4 DEG C of refrigerators and are dried；
(3) electrode obtained in step (2) is cleaned with PBS, continues in 6 ~ 10 μ L concentration of electrode surface drop coating to be 20 μ g/
The alkaline phosphatase enzyme solutions of mL are saved in 4 DEG C of refrigerators and are dried；
(4) electrode obtained in step (3) is cleaned with PBS, continues in 8 ~ 12 μ L concentration of electrode surface drop coating to be 100 μ g/
The bovine serum albumin solution of mL is saved in 4 DEG C of refrigerators and is dried；
(5) electrode obtained in step (4) is cleaned with PBS, is saved in 4 DEG C of refrigerators after drying, obtains optical electro-chemistry
Salbutamol sensor；
The Fe-TiO2/MoS2Colloidal sol is by the Fe-TiO of 50 mg2/MoS2Powder is dissolved in 10 mL ultrapure waters, and ultrasound
The hydrosol obtained after 30 min；
The PBS is the phosphate buffer solution of 10 mmol/L, and the pH value of the phosphate buffer solution is 7.4.
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