CN107478698B - A kind of preparation method and application of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor - Google Patents
A kind of preparation method and application of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor Download PDFInfo
- Publication number
- CN107478698B CN107478698B CN201710655557.2A CN201710655557A CN107478698B CN 107478698 B CN107478698 B CN 107478698B CN 201710655557 A CN201710655557 A CN 201710655557A CN 107478698 B CN107478698 B CN 107478698B
- Authority
- CN
- China
- Prior art keywords
- aflatoxin
- preparation
- solution
- octahedron
- silver sulfide
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Nanotechnology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to immunoassays and biosensor technique field, provide a kind of preparation method and application of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor.Using WO3For substrate, excellent electric conductivity and big surface area can effectively reduce background signal.Using silver ion functionalization octahedron pucherite nanoparticle label aflatoxin, by the narrow band gap Ag that vulcanized sodium in-situ preparation high optoelectronic conversion ratio is added dropwise2S generates photo-signal as signal amplified material, by the way that the LED light irradiation of visible wavelength is lower.Carrier octahedron pucherite and silver sulfide band-gap degree are good, can be further improved the photoelectric conversion efficiency of silver sulfide, realize the detection of common aflatoxin, with high specificity, high sensitivity, detection limit is low, has important scientific meaning and application value to the detection of aflatoxin.
Description
Technical field
The invention belongs to novel function nanometer material, immunoassay and biosensor technique fields, provide a kind of original position
Generate the preparation method and application of silver sulfide competitive type aflatoxin optical electro-chemistry sensor.
Background technique
In recent years, food pollution is on the rise and frequently, not only causes huge economic loss, can also seriously affect the mankind
Health.Aflatoxin is to be grown in grain (wheat, corn, barley, oat, rye, rice and broomcorn millet class etc.), flower by a variety of
What aspergillus and mould on the crops such as life, vegetables generated.Due to widely distributed, it is easy to pass through the cereal of pollution or feed etc.
Into food chain, and then the animal-derived foods such as milk, egg, meat are polluted, indirectly entered in human body, ultimately causes lesions of liver and kidney, numerous
Grow the serious consequences such as obstacle, immunosupress, carcinogenic teratogenesis.
Food inspection is to guarantee the important link of food safety, and the present invention provides a kind of quick, easy, sensitivity and choosings
The high optical electro-chemistry immunoassay method of selecting property.Optical electro-chemistry sensor is the photoelectric conversion based on substance to determine that determinand is dense
A kind of detection device of degree, optical electro-chemistry detection method have the characteristics that high sensitivity, equipment are simple, are easy to be miniaturized,
As a kind of analysis method of great application potential, have broad application prospects in food, environment, medicine and other fields.
The invention patent successfully constructs the optical electro-chemistry immunosensor that aflatoxin is detected under excited by visible light.
The sensor is with WO3For substrate, excellent electric conductivity and big surface area can effectively reduce background signal.Utilize silver ion function
Energyization octahedron pucherite nanoparticle label aflatoxin, by the way that the narrow of vulcanized sodium in-situ preparation high optoelectronic conversion ratio is added dropwise
Band gap silver sulfide is as signal amplified material.It is prepared by the present invention to be sensed based on in-situ preparation silver sulfide competitive type optical electro-chemistry
Device has many advantages, such as that inexpensive, highly sensitive, specificity is good, detection is quick, preparation process is simple, realizes in visible light region
To quick, the Sensitive Detection of aflatoxin, the deficiency of current Determination Methods of Aflatoxins is effectively overcome.
Summary of the invention
The present invention provides a kind of preparation sides of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor
Method and application realize the super sensitivity detection to aflatoxin.
An object of the present invention is to provide a kind of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensing
The preparation method of device.
The second object of the present invention is to pass prepared in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry
Sensor, for detecting aflatoxin.
Technical solution of the present invention includes the following steps.
A kind of preparation method of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor, including it is following several
A step:
(1) ITO electro-conductive glass is cut to 2.5cm × 1.0cm size, is successively cleaned with acetone, ethyl alcohol and ultrapure water
30min is dried with nitrogen；
(2) WO of 10 μ L, 3 ~ 5 mg/mL are taken3It is added drop-wise to the conducting surface of electro-conductive glass, is dried at room temperature, 300 DEG C are forged
120 min are burnt, room temperature is cooled to, in WO3The thioacetic acid of 3 ~ 5 μ L, 0.1mol/L is added dropwise in modified electrode surface, dries in the air at room temperature
It is dry, the EDC/NHS mixed liquor of 3 ~ 5 μ L, dry, ultrapure water in 4 DEG C of refrigerators is added dropwise；
(3) the aflatoxin antigen of 4 ~ 6 μ L, 10 μ g/mL are added drop-wise to electrode surface, drying in 4 DEG C of refrigerators surpasses
Pure water rinsing；
(4) continue 2 ~ 4 μ L, the BSA solution that mass fraction is 1% being added drop-wise to electrode surface, with enclosed-electrode surface
Upper nonspecific activity site is dried in 4 DEG C of refrigerators, ultrapure water electrode surface；
(5) 5 μ L, the ng/mL of 10 pg/mL ~ 60 aflatoxin and silver ion functionalization octahedron bismuth vanadate yellow is bent
The mixed liquor of mould toxin antibody marker is added drop-wise to electrode surface, hatches 30min in 4 DEG C of refrigerators, continues that 3 μ L, 0.08 are added dropwise
The Na of mol/L2S dries in 4 DEG C of refrigerators, ultrapure water electrode surface, and it is yellow that a kind of in-situ preparation silver sulfide competitive type is made
Aspertoxin optical electro-chemistry sensor；
The EDS/NHS mixed liquor refers to containing 1 × 10-21- ethyl -3- (3- dimethyl aminopropyl) carbon two of mol/L
Imines and 2 × 10-3The mixed aqueous solution of the n-hydroxysuccinimide of mol/L.
WO3Preparation, steps are as follows:
It takes 10 ~ 12 mmol sodium tungstates to be dissolved in 20 mL ultrapure waters, pH is adjusted to 1.5 with the hydrochloric acid of 4mol/L, is added
Enter 20 ~ 24 mmol sodium chloride and 10 ~ 12 mmol oxalic acid, after stirring 30 min, adds 50 mL ultrapure waters, then turn
It moves on in polytetrafluoroethylene (PTFE) autoclave, 2 ~ 3 h is reacted at 150 DEG C, are cooled to room temperature, be centrifugated, respectively with ultrapure
WO is made in water and dehydrated alcohol centrifuge washing, 80 DEG C of 12 h of vacuum drying3Powder.
The preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution, steps are as follows:
(1) preparation of octahedra pucherite
It takes the bismuth nitrate of 3.6mmol to be dissolved in 30 ~ 35mL, 2.4 mol/L nitric acid, forms solution A；Take the inclined vanadium of 3.6mmol
Sour ammonium is dissolved in 30 ~ 35 mL, 1.9 mol/L sodium hydroxides, forms solution B；0.6 g 12 is separately added into two solution of A, B
Sodium alkyl benzene sulfonate stirs 30 min respectively, and two solution of A, B is successively transferred in polytetrafluoroethylene (PTFE) autoclave and is mixed, and 200 DEG C
3 ~ 5 h of lower reaction, are cooled to room temperature, centrifuge separation, three times with ultrapure water and dehydrated alcohol centrifuge washing, 80 DEG C of vacuum drying
18 ~ 22 h obtain octahedra pucherite；
(2) preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution
Weigh the thioacetic acid that 15 ~ 25 mg octahedra pucherite obtained above is dissolved in 2 mL, 0.1mol/L, ultrasound
20 min, are added the silver nitrate of 7.5 ~ 8.5 mL, 20mmol/L, and room-temperature water bath dark place vibrates 12 h；It is added 500 μ L's
EDC/NHS mixed liquor vibrates 2 h；Centrifuge washing is dispersed with the buffer solution of the PBS of pH 7.0, and it is yellow that 1mL, 10 μ g/mL are added
Aspertoxin antibody vibrates 24 h；It is molten to be dispersed in 2 mL, the PBS containing the pH 7.0 that mass fraction is 1.0% BSA for centrifuge washing
In liquid, silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution is made, storage in 4 DEG C of refrigerators in case
With.
4. the ng/mL of 10 pg/mL ~ 60 aflatoxin and silver ion functionalization octahedron bismuth vanadate yellow aspergillus poison
The preparation of the mixed liquor of plain antibody marker, steps are as follows:
By the silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker of 1mL, 2mg/mL and 10 pg/mL ~
The aflatoxin standard solution of 60 ng/mL mixes, and 60 min is vibrated in 4 DEG C of constant-temperature shaking incubators, with the PBS of pH 7.0
Centrifuge washing is dispersed in the PBS buffer solution of the pH 7.0 of 1mL, be made the ng/mL of 10 pg/mL ~ 60 aflatoxin with
The mixed liquor of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker.
The detection of aflatoxin, steps are as follows:
(1) it is tested using electrochemical workstation with three-electrode system, prepared sensor is working electrode, saturation
Calomel electrode is reference electrode, and platinum electrode is auxiliary electrode, in the PBS containing 0.12 mol/L ascorbic acid of 10mL, pH7.0
Buffer solution is tested；
(2) used time m- current method detects analyte, and setting voltage is 0.1V, runing time 100s, irradiation LED
Lamp wavelength is 400 ~ 450 nm；
(3) it after background current tends towards stability, turns on light 20 s of prolonged exposure every 20 s, then records photoelectric current, draw
Working curve；
(4) aflatoxin standard solution is replaced to detect aflatoxin sample solution to be measured, the knot of detection
Fruit can be checked in by working curve.
The aflatoxin, selected from one of following: aflatoxin B1, aflatoxin B 2, aflatoxin G 1,
Aflatoxin G 2.
Raw materials of the present invention can be bought in chemical reagents corporation or biopharmaceutical company.
Beneficial achievement of the invention
(1) present invention uses with semiconductor WO3For substrate, excellent electric conductivity and big surface area can be effectively reduced
Background signal is crosslinking with thioacetic acid, 1- ethyl -3- (3- dimethyl aminopropyl) carbodiimide/n-hydroxysuccinimide
Agent promotes effective combination of antibody.
(2) Na is added dropwise as competition marker using silver ion functionalization octahedron pucherite on it2S solution, it is in situ
Silver sulfide competitive type aflatoxin optical electro-chemistry sensor is generated, quick, the sensitive detection to aflatoxin is realized；
(3) detection of a kind of in-situ preparation silver sulfide competitive type optical electro-chemistry sensor to aflatoxin, operation letter
Single, signal response range is wide, and to aflatoxin B1, aflatoxin B 2, the range of linearity of detection is 10 pg/mL ~ 50
Ng/mL, detection are limited to 3.3pg/mL to aflatoxin G 1, aflatoxin G 2,10 pg/mL ~ 60 of the range of linearity of detection
Ng/mL, detection are limited to 3.3 pg/mL, show that this competitive type optical electro-chemistry sensor can achieve the purpose of Accurate Determining；
(4) semiconductor WO3It is excellent to possess good electric conductivity, photoelectric activity, big surface area, high stability and low cost etc.
Point.Octahedra pucherite, not only promotes the absorption of visible light, and can accelerate the separation of electron hole pair and increase its conduction
Property.In silver ion functionalization octahedron pucherite in-situ preparation silver sulfide, the good competitive type photoelectric transfer of band-gap is obtained
Sensor, to improve incident photon-to-electron conversion efficiency and realize highly sensitive detection.
Specific embodiment
Now the present invention is further illustrated by specific embodiment, but not limited to this
A kind of preparation method of the in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor of embodiment 1, packet
Include following steps:
(1) ITO electro-conductive glass is cut to 2.5cm × 1.0cm size, is successively cleaned with acetone, ethyl alcohol and ultrapure water
30min is dried with nitrogen；
(2) WO of 10 μ L, 3 mg/mL are taken3It is added drop-wise to the conducting surface of electro-conductive glass, is dried at room temperature, 300 DEG C of calcinings 120
Min is cooled to room temperature, in WO3The thioacetic acid of 3 μ L, 0.1mol/L is added dropwise in modified electrode surface, dries at room temperature, is added dropwise 3
The EDC/NHS mixed liquor of μ L, dry in 4 DEG C of refrigerators, ultrapure water；
(3) the aflatoxin antigen of 4 μ L, 10 μ g/mL are added drop-wise to electrode surface, drying in 4 DEG C of refrigerators is ultrapure
Water rinses；
(4) continue 2 μ L, the BSA solution that mass fraction is 1% being added drop-wise to electrode surface, with non-on enclosed-electrode surface
Activity specific site is dried in 4 DEG C of refrigerators, ultrapure water electrode surface；
(5) 5 μ L, the ng/mL of 10 pg/mL ~ 60 aflatoxin and silver ion functionalization octahedron bismuth vanadate yellow is bent
The mixed liquor of mould toxin antibody marker is added drop-wise to electrode surface, hatches 30min in 4 DEG C of refrigerators, continues that 3 μ L, 0.08 are added dropwise
The Na of mol/L2S dries in 4 DEG C of refrigerators, ultrapure water electrode surface, and it is yellow that a kind of in-situ preparation silver sulfide competitive type is made
Aspertoxin optical electro-chemistry sensor；
The EDS/NHS mixed liquor refers to containing 1 × 10-21- ethyl -3- (3- dimethyl aminopropyl) carbon two of mol/L
Imines and 2 × 10-3The mixed aqueous solution of the n-hydroxysuccinimide of mol/L.
A kind of preparation method of the in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor of embodiment 2, packet
Include following steps:
(1) ITO electro-conductive glass is cut to 2.5cm × 1.0cm size, is successively cleaned with acetone, ethyl alcohol and ultrapure water
30min is dried with nitrogen；
(2) WO of 10 μ L, 4mg/mL are taken3It is added drop-wise to the conducting surface of electro-conductive glass, is dried at room temperature, 300 DEG C of calcinings 120
Min is cooled to room temperature, in WO3The thioacetic acid of 4 μ L, 0.1mol/L is added dropwise in modified electrode surface, dries at room temperature, is added dropwise 4
The EDC/NHS mixed liquor of μ L, dry in 4 DEG C of refrigerators, ultrapure water；
(3) the aflatoxin antigen of 5 μ L, 10 μ g/mL are added drop-wise to electrode surface, drying in 4 DEG C of refrigerators is ultrapure
Water rinses；
(4) continue 3 μ L, the BSA solution that mass fraction is 1% being added drop-wise to electrode surface, with non-on enclosed-electrode surface
Activity specific site is dried in 4 DEG C of refrigerators, ultrapure water electrode surface；
(5) 5 μ L, the ng/mL of 10 pg/mL ~ 60 aflatoxin and silver ion functionalization octahedron bismuth vanadate yellow is bent
The mixed liquor of mould toxin antibody marker is added drop-wise to electrode surface, hatches 30min in 4 DEG C of refrigerators, continues that 3 μ L, 0.08 are added dropwise
The Na of mol/L2S dries in 4 DEG C of refrigerators, ultrapure water electrode surface, and it is yellow that a kind of in-situ preparation silver sulfide competitive type is made
Aspertoxin optical electro-chemistry sensor；
The EDS/NHS mixed liquor refers to containing 1 × 10-21- ethyl -3- (3- dimethyl aminopropyl) carbon two of mol/L
Imines and 2 × 10-3The mixed aqueous solution of the n-hydroxysuccinimide of mol/L.
A kind of preparation method of the in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor of embodiment 3, packet
Include following steps:
(1) ITO electro-conductive glass is cut to 2.5cm × 1.0cm size, is successively cleaned with acetone, ethyl alcohol and ultrapure water
30min is dried with nitrogen；
(2) WO of 10 μ L, 5 mg/mL are taken3It is added drop-wise to the conducting surface of electro-conductive glass, is dried at room temperature, 300 DEG C of calcinings 120
Min is cooled to room temperature, in WO3The thioacetic acid of 5 μ L, 0.1mol/L is added dropwise in modified electrode surface, dries at room temperature, and 5 μ are added dropwise
The EDC/NHS mixed liquor of L, dry in 4 DEG C of refrigerators, ultrapure water；
(3) the aflatoxin antigen of 6 μ L, 10 μ g/mL are added drop-wise to electrode surface, drying in 4 DEG C of refrigerators is ultrapure
Water rinses；
(4) continue 4 μ L, the BSA solution that mass fraction is 1% being added drop-wise to electrode surface, with non-on enclosed-electrode surface
Activity specific site is dried in 4 DEG C of refrigerators, ultrapure water electrode surface；
(5) 5 μ L, the ng/mL of 10 pg/mL ~ 60 aflatoxin and silver ion functionalization octahedron bismuth vanadate yellow is bent
The mixed liquor of mould toxin antibody marker is added drop-wise to electrode surface, hatches 30min in 4 DEG C of refrigerators, continues that 3 μ L, 0.08 are added dropwise
The Na of mol/L2S dries in 4 DEG C of refrigerators, ultrapure water electrode surface, and it is yellow that a kind of in-situ preparation silver sulfide competitive type is made
Aspertoxin optical electro-chemistry sensor；
The EDS/NHS mixed liquor refers to containing 1 × 10-21- ethyl -3- (3- dimethyl aminopropyl) carbon two of mol/L
Imines and 2 × 10-3The mixed aqueous solution of the n-hydroxysuccinimide of mol/L.
WO described in embodiment 43Preparation, steps are as follows:
It takes 10 mmol sodium tungstates to be dissolved in 20 mL ultrapure waters, pH is adjusted to 1.5 with the hydrochloric acid of 4mol/L, is added 20
Mmol sodium chloride and 10 mmol oxalic acid add 50 mL ultrapure waters, are then transferred into polytetrafluoroethylene (PTFE) height after stirring 30 min
It presses in reaction kettle, 2 h is reacted at 150 DEG C, are cooled to room temperature, be centrifugated, washed respectively with ultrapure water and dehydrated alcohol centrifugation
It washs, 80 DEG C of 12 h of vacuum drying, WO is made3Powder.
WO described in embodiment 53Preparation, steps are as follows:
It takes 11 mmol sodium tungstates to be dissolved in 20 mL ultrapure waters, pH is adjusted to 1.5 with the hydrochloric acid of 4mol/L, is added 22
Mmol sodium chloride and 11 mmol oxalic acid add 50 mL ultrapure waters, are then transferred into polytetrafluoroethylene (PTFE) height after stirring 30 min
It presses in reaction kettle, 2.5 h is reacted at 150 DEG C, are cooled to room temperature, be centrifugated, washed respectively with ultrapure water and dehydrated alcohol centrifugation
It washs, 80 DEG C of 12 h of vacuum drying, WO is made3Powder.
WO described in embodiment 63Preparation, steps are as follows:
It takes 12 mmol sodium tungstates to be dissolved in 20 mL ultrapure waters, pH is adjusted to 1.5 with the hydrochloric acid of 4mol/L, is added 24
Mmol sodium chloride and 12 mmol oxalic acid add 50 mL ultrapure waters, are then transferred into polytetrafluoroethylene (PTFE) height after stirring 30 min
It presses in reaction kettle, 3 h is reacted at 150 DEG C, are cooled to room temperature, be centrifugated, washed respectively with ultrapure water and dehydrated alcohol centrifugation
It washs, 80 DEG C of 12 h of vacuum drying, WO is made3Powder.
The preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution described in embodiment 7.,
Steps are as follows:
(1) preparation of octahedra pucherite
It takes the bismuth nitrate of 3.6mmol to be dissolved in 30 mL, 2.4 mol/L nitric acid, forms solution A；Take 3.6mmol ammonium metavanadate
It is dissolved in 30mL, 1.9 mol/L sodium hydroxides, forms solution B；0.6 g dodecyl benzene sulfonic acid is separately added into two solution of A, B
Sodium stirs 30 min respectively, and two solution of A, B is successively transferred in polytetrafluoroethylene (PTFE) autoclave and is mixed, 3 h are reacted at 200 DEG C,
It is cooled to room temperature, is centrifugated, three times with ultrapure water and dehydrated alcohol centrifuge washing, 80 DEG C of 18 h of vacuum drying obtain octahedral
Body pucherite；
(2) preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution
Weigh the thioacetic acid that 15 mg octahedra pucherite obtained above is dissolved in 2 mL, 0.1mol/L, ultrasound 20
Min, is added the silver nitrate of 7.5 mL, 20mmol/L, and room-temperature water bath dark place vibrates 12 h；The EDC/NHS mixing of 500 μ L is added
Liquid vibrates 2 h；Centrifuge washing is dispersed with the buffer solution of the PBS of pH 7.0, it is anti-that 1mL, 10 μ g/mL aflatoxin is added
24 h of oscillation body；Centrifuge washing, be dispersed in 2 mL, containing mass fraction be 1.0% BSA pH 7.0 PBS solution in, be made silver
Ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution, storage is with spare in 4 DEG C of refrigerators.
The preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution described in embodiment 8.,
Steps are as follows:
(1) preparation of octahedra pucherite
It takes the bismuth nitrate of 3.6mmol to be dissolved in 32.5mL, 2.4 mol/L nitric acid, forms solution A；Take 3.6mmol ammonium metavanadate
32.5 mL, 1.9 mol/L sodium hydroxides are dissolved in, solution B is formed；0.6 g detergent alkylate is separately added into two solution of A, B
Sodium sulfonate stirs 30 min respectively, and two solution of A, B is successively transferred in polytetrafluoroethylene (PTFE) autoclave and is mixed, is reacted at 200 DEG C
4 h, are cooled to room temperature, and centrifuge separation, three times with ultrapure water and dehydrated alcohol centrifuge washing, 80 DEG C of 20 h of vacuum drying are obtained
Octahedra pucherite；
(2) preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution
Weigh the thioacetic acid that 20 mg octahedra pucherite obtained above is dissolved in 2 mL, 0.1mol/L, ultrasound 20
Min, is added the silver nitrate of 8.0 mL, 20mmol/L, and room-temperature water bath dark place vibrates 12 h；The EDC/NHS mixing of 500 μ L is added
Liquid vibrates 2 h；Centrifuge washing is dispersed with the buffer solution of the PBS of pH 7.0, it is anti-that 1mL, 10 μ g/mL aflatoxin is added
24 h of oscillation body；Centrifuge washing, be dispersed in 2 mL, containing mass fraction be 1.0% BSA pH 7.0 PBS solution in, be made silver
Ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution, storage is with spare in 4 DEG C of refrigerators.
The preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution described in embodiment 9.,
Steps are as follows:
(1) preparation of octahedra pucherite
It takes the bismuth nitrate of 3.6mmol to be dissolved in 35 mL, 2.4 mol/L nitric acid, forms solution A；Take 3.6mmol ammonium metavanadate
35 mL, 1.9 mol/L sodium hydroxides are dissolved in, solution B is formed；0.6 g detergent alkylate sulphur is separately added into two solution of A, B
Sour sodium stirs 30 min respectively, and two solution of A, B is successively transferred in polytetrafluoroethylene (PTFE) autoclave and is mixed, reacts 5 at 200 DEG C
H is cooled to room temperature, and centrifuge separation, three times with ultrapure water and dehydrated alcohol centrifuge washing, 80 DEG C of 22 h of vacuum drying obtain eight
Face body pucherite；
(2) preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution
Weigh the thioacetic acid that 25 mg octahedra pucherite obtained above is dissolved in 2 mL, 0.1mol/L, ultrasound 20
Min, is added the silver nitrate of 8.5 mL, 20mmol/L, and room-temperature water bath dark place vibrates 12 h；The EDC/NHS mixing of 500 μ L is added
Liquid vibrates 2 h；Centrifuge washing is dispersed with the buffer solution of the PBS of pH 7.0, it is anti-that 1mL, 10 μ g/mL aflatoxin is added
24 h of oscillation body；Centrifuge washing, be dispersed in 2 mL, containing mass fraction be 1.0% BSA pH 7.0 PBS solution in, be made silver
Ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution, storage is with spare in 4 DEG C of refrigerators.
The ng/mL of 10 pg/mL ~ 60 aflatoxin described in embodiment 10. and silver ion functionalization octahedron pucherite
The preparation of the mixed liquor of aflatoxin antibody marker, steps are as follows:
By the silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker of 1mL, 2mg/mL and 10 pg/mL ~
The aflatoxin standard solution of 60 ng/mL mixes, and 60 min is vibrated in 4 DEG C of constant-temperature shaking incubators, with the PBS of pH 7.0
Centrifuge washing is dispersed in the PBS buffer solution of the pH 7.0 of 1mL, be made the ng/mL of 10 pg/mL ~ 60 aflatoxin with
The mixed liquor of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker.
The detection of 11. aflatoxin B1 of embodiment
(1) it is tested using electrochemical workstation with three-electrode system, prepared sensor is working electrode, saturation
Calomel electrode is reference electrode, and platinum electrode is auxiliary electrode, in the PBS containing 0.12 mol/L ascorbic acid of 10mL, pH7.0
Buffer solution is tested；
(2) used time m- current method detects analyte, and setting voltage is 0.1V, runing time 100s, irradiation LED
Lamp wavelength is 400 ~ 450 nm；
(3) it after background current tends towards stability, turns on light 20 s of prolonged exposure every 20 s, then records photoelectric current, draw
Working curve；
(4) range of linearity for measuring aflatoxin B1 in sample is the ng/mL of 10 pg/mL ~ 50, and detection is limited to 3.3
pg/mL。
The detection of 12. aflatoxin B 2 of embodiment
Aflatoxin B 2 in sample is detected according to the method for embodiment 11, the range of linearity be 10 pg/mL ~
50 ng/mL, detection are limited to 3.3 pg/mL.
The detection of 13. aflatoxin G 1 of embodiment
Aflatoxin G 1 in sample is detected according to the method for embodiment 11, the range of linearity be 10 pg/mL ~
60 ng/mL, detection are limited to 3.3 pg/mL.
The detection of 14. aflatoxin G 2 of embodiment
Aflatoxin G 2 in sample is detected according to the method for embodiment 11, the range of linearity be 10 pg/mL ~
60 ng/mL, detection are limited to 3.3 pg/mL.
Claims (7)
1. a kind of preparation method of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor, which is characterized in that
Including the following steps:
(1) ITO electro-conductive glass is cut to 2.5cm × 1.0cm size, is successively cleaned with acetone, ethyl alcohol and ultrapure water
30min is dried with nitrogen；
(2) WO of 10 μ L, 3 ~ 5 mg/mL are taken3It is added drop-wise to the conducting surface of electro-conductive glass, is dried at room temperature, 300 DEG C of calcinings 120
Min is cooled to room temperature, in WO3The thioacetic acid of 3 ~ 5 μ L, 0.1mol/L is added dropwise in modified electrode surface, dries at room temperature, drips
Add the EDC/NHS mixed liquor of 3 ~ 5 μ L, dry, ultrapure water in 4 DEG C of refrigerators；
(3) the aflatoxin antigen of 4 ~ 6 μ L, 10 μ g/mL are added drop-wise to electrode surface, dry, ultrapure water in 4 DEG C of refrigerators
It rinses；
(4) continue 2 ~ 4 μ L, the BSA solution that mass fraction is 1% being added drop-wise to electrode surface, with non-spy on enclosed-electrode surface
Specific activities site is dried in 4 DEG C of refrigerators, ultrapure water electrode surface；
(5) by 5 μ L, the ng/mL of 10 pg/mL ~ 60 aflatoxin and silver ion functionalization octahedron bismuth vanadate yellow aspertoxin
The mixed liquor of antibody marker is added drop-wise to electrode surface, hatch 30min in 4 DEG C of refrigerators, continues that 3 μ L, 0.08mol/L are added dropwise
Na2S dries in 4 DEG C of refrigerators, and a kind of in-situ preparation silver sulfide competitive type aflatoxin is made in ultrapure water electrode surface
Optical electro-chemistry sensor；
The EDS/NHS mixed liquor refers to containing 1 × 10-21- ethyl -3- (3- dimethyl aminopropyl) carbodiimide of mol/L
With 2 × 10-3The mixed aqueous solution of the n-hydroxysuccinimide of mol/L.
2. a kind of preparation of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor as described in claim 1
Method, which is characterized in that the WO3Preparation, steps are as follows:
It takes 10 ~ 12 mmol sodium tungstates to be dissolved in 20 mL ultrapure waters, pH is adjusted to 1.5 with the hydrochloric acid of 4mol/L, is added 20
~ 24 mmol sodium chloride and 10 ~ 12 mmol oxalic acid add 50 mL ultrapure waters, are then transferred into poly- after stirring 30 min
In tetrafluoroethene autoclave, 2 ~ 3h is reacted at 150 DEG C, is cooled to room temperature, be centrifugated, respectively with ultrapure water and anhydrous
WO is made in ethyl alcohol centrifuge washing, 80 DEG C of 12 h of vacuum drying3Powder.
3. a kind of preparation of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor as described in claim 1
Method, which is characterized in that the preparation of the silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution, packet
Include following steps:
(1) preparation of octahedra pucherite
It takes the bismuth nitrate of 3.6mmol to be dissolved in 30 ~ 35mL, 2.4 mol/L nitric acid, forms solution A；Take 3.6mmol ammonium metavanadate molten
In 30 ~ 35 mL, 1.9 mol/L sodium hydroxides, solution B is formed；0.6 g detergent alkylate is separately added into two solution of A, B
Sodium sulfonate stirs 30 min respectively, and two solution of A, B is successively transferred in polytetrafluoroethylene (PTFE) autoclave and is mixed, is reacted at 200 DEG C
3 ~ 5 h, are cooled to room temperature, centrifuge separation, three times with ultrapure water and dehydrated alcohol centrifuge washing, 80 DEG C of vacuum drying 18 ~ 22
H obtains octahedra pucherite；
(2) preparation of silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution
Weigh the thioacetic acid that 15 ~ 25mg octahedra pucherite obtained above is dissolved in 2 mL, 0.1mol/L, ultrasound 20
Min, is added the silver nitrate of 7.5 ~ 8.5 mL, 20mmol/L, and room-temperature water bath dark place vibrates 12h；The EDC/NHS of 500 μ L is added
Mixed liquor vibrates 2h；Centrifuge washing is dispersed with the buffer solution of the PBS of pH 7.0,1mL, 10 μ g/mL aflatoxin is added
Antibody vibrates for 24 hours；Centrifuge washing, be dispersed in 2 mL, containing mass fraction be 1.0% BSA pH 7.0 PBS solution in, be made
Silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker solution, storage is with spare in 4 DEG C of refrigerators.
4. a kind of preparation of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor as described in claim 1
Method, which is characterized in that the ng/mL of 10 pg/mL ~ 60 aflatoxin and silver ion functionalization octahedron bismuth vanadate yellow
The preparation of the mixed liquor of aspertoxin antibody marker, steps are as follows:
By the silver ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker of 1mL, 2mg/mL and 10 pg/mL ~ 60
The aflatoxin standard solution of ng/mL mixes, and vibrates 60 min in 4 DEG C of constant-temperature shaking incubators, with the PBS of pH 7.0 from
Heart washing, is dispersed in the PBS buffer solution of the pH 7.0 of 1mL, and the ng/mL of 10 pg/mL ~ 60 aflatoxin and silver is made
The mixed liquor of ion functionalization octahedron bismuth vanadate yellow aspertoxin antibody marker.
5. such as a kind of described in any item in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry biographies of claim 1 ~ 4
The preparation method of sensor, which is characterized in that the aflatoxin is selected from one of following: aflatoxin B1, aflatoxin
B2, aflatoxin G 1, aflatoxin G 2.
6. a kind of in-situ preparation silver sulfide competitive type aflatoxin of preparation method preparation as described in claim 1 is photoelectrochemical
Sensor is learned, for the detection of aflatoxin, detecting step is as follows:
(1) it is tested using electrochemical workstation with three-electrode system, prepared sensor is working electrode, is saturated calomel
Electrode is reference electrode, and platinum electrode is auxiliary electrode, in the PBS buffering of the ascorbic acid containing 0.12mol/L of 10mL, pH7.0
Solution is tested,
(2) used time m- current method detects analyte, and setting voltage is 0.1V, runing time 100s, irradiation LED lamp wave
A length of 400 ~ 450 nm,
(3) it after background current tends towards stability, turns on light 20 s of prolonged exposure every 20s, then records photoelectric current, it is bent to draw work
Line；
(4) aflatoxin standard solution is replaced to detect aflatoxin sample solution to be measured, the result of detection can
It is checked in by working curve.
7. a kind of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor as claimed in claim 6, feature
It is, the aflatoxin is selected from one of following: aflatoxin B1, aflatoxin B 2, aflatoxin G 1, aspergillus flavus
Toxin G2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710655557.2A CN107478698B (en) | 2017-08-03 | 2017-08-03 | A kind of preparation method and application of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710655557.2A CN107478698B (en) | 2017-08-03 | 2017-08-03 | A kind of preparation method and application of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107478698A CN107478698A (en) | 2017-12-15 |
| CN107478698B true CN107478698B (en) | 2019-08-20 |
Family
ID=60598080
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710655557.2A Expired - Fee Related CN107478698B (en) | 2017-08-03 | 2017-08-03 | A kind of preparation method and application of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107478698B (en) |
Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108828036B (en) * | 2018-06-12 | 2020-09-22 | 湖北大学 | Molecular imprinting photoelectrochemical sensor based on graphene oxide and cadmium sulfide quantum dot heterostructure and preparation method and application thereof |
| CN108845015B (en) * | 2018-06-28 | 2020-09-01 | 济南大学 | Preparation method and application of photoelectrochemical aflatoxin B1 sensor based on tungsten trioxide composite material |
| CN109142745B (en) * | 2018-07-27 | 2021-08-20 | 济南大学 | Preparation method and application of photoelectrochemical immunosensor based on stannic oxide/cadmium carbonate/cadmium sulfide |
| CN109746001A (en) * | 2018-12-13 | 2019-05-14 | 广东工业大学 | A kind of tin oxide photonic crystal load tungsten oxide and vulcanization silver composite membrane and its preparation method and application |
| CN109926068B (en) * | 2019-03-08 | 2020-08-07 | 武汉大学 | Silver sulfide/tungsten trioxide-based Z-type photocatalytic material, and preparation method and application thereof |
| CN110297023B (en) * | 2019-07-09 | 2021-05-28 | 济南大学 | Preparation method and application of self-enhanced photoelectrochemical immunosensor for detecting procalcitonin electrochemical catalysis assistance |
| CN110243892A (en) * | 2019-07-29 | 2019-09-17 | 济南大学 | A kind of preparation method of the optical electro-chemistry Procalcitonin biosensor based on flower-shaped tungstic acid composite material |
| CN111220671A (en) * | 2020-03-30 | 2020-06-02 | 济南大学 | Detect aflatoxin B1Preparation method and application of electrochemiluminescence sensing electrode |
| CN112986346B (en) * | 2021-02-24 | 2022-11-11 | 青岛科技大学 | Based on BiVO 4 /Ag 2 S heterojunction photoelectric chemical biosensor and application thereof |
| CN113637943B (en) * | 2021-07-28 | 2023-04-25 | 杭州电子科技大学 | Preparation method of photosensitive carbon disulfide sensor |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013244440A (en) * | 2012-05-24 | 2013-12-09 | Sharp Corp | Water purifying apparatus |
| US8702962B1 (en) * | 2007-05-25 | 2014-04-22 | The United States Of America As Represented By The Administrator Of National Aeronautics And Space Administration | Carbon dioxide gas sensors and method of manufacturing and using same |
| CN106198501A (en) * | 2016-07-05 | 2016-12-07 | 济南大学 | A kind of preparation method of the Electrochemiluminescsensor sensor detecting aflatoxin |
| CN106807409A (en) * | 2017-03-14 | 2017-06-09 | 华东理工大学 | A kind of preparation method of Z-type catalysis material vanadic acid bismuth silver silver bromide |
| CN106855533A (en) * | 2017-01-17 | 2017-06-16 | 济南大学 | One kind is based on rGO/CdSe:The preparation method of the PSA interlayer type Optical Electro-Chemistry sensor of Ca |
-
2017
- 2017-08-03 CN CN201710655557.2A patent/CN107478698B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8702962B1 (en) * | 2007-05-25 | 2014-04-22 | The United States Of America As Represented By The Administrator Of National Aeronautics And Space Administration | Carbon dioxide gas sensors and method of manufacturing and using same |
| JP2013244440A (en) * | 2012-05-24 | 2013-12-09 | Sharp Corp | Water purifying apparatus |
| CN106198501A (en) * | 2016-07-05 | 2016-12-07 | 济南大学 | A kind of preparation method of the Electrochemiluminescsensor sensor detecting aflatoxin |
| CN106855533A (en) * | 2017-01-17 | 2017-06-16 | 济南大学 | One kind is based on rGO/CdSe:The preparation method of the PSA interlayer type Optical Electro-Chemistry sensor of Ca |
| CN106807409A (en) * | 2017-03-14 | 2017-06-09 | 华东理工大学 | A kind of preparation method of Z-type catalysis material vanadic acid bismuth silver silver bromide |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107478698A (en) | 2017-12-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107478698B (en) | A kind of preparation method and application of in-situ preparation silver sulfide competitive type aflatoxin optical electro-chemistry sensor | |
| US10913892B1 (en) | Method for preparing ratiometric fluorescent probe for melamine based on silver nanocluster complex | |
| Liu et al. | Recent progress on electrochemical biosensing of aflatoxins: A review | |
| CN104297464B (en) | A kind of in-situ preparation CdS mycotoxin Optical Electro-Chemistry transducer production method and application | |
| CN106802315A (en) | A kind of preparation method and application of ochratoxin A Optical Electro-Chemistry sensor | |
| CN109060905A (en) | The preparation method of additive Mn cadmium selenide enhancing bismuth tungstate-cadmium sulfide amyloid beta competitive type optical electro-chemistry sensor | |
| CN108120750B (en) | A kind of preparation method and application of zearalanol optical electro-chemistry sensor | |
| CN104297305A (en) | Making method and application of CdS sensitized TiO2 environmental estrogen photoelectrochemical sensor | |
| CN107831198A (en) | A kind of preparation method and application of the optical electro-chemistry cTnI sensor based on multistage micron cube zinc stannate composite | |
| Xu et al. | Using SiO2/PDA-Ag NPs to dual-inhibited photoelectrochemical activity of CeO2-CdS composites fabricated a novel immunosensor for BNP ultrasensitive detection | |
| CN108845015A (en) | A kind of preparation method and application of the optical electro-chemistry aflatoxin B1 sensor based on tungstic acid composite material | |
| CN104849331A (en) | Preparing method of photoelectrochemical sensor based on sandwich cardiac troponin T marked by Ag2Se@CdSe and application | |
| CN105259236A (en) | Preparation method of photoelectrochemical sensor for detecting dexamethasone based on in-situ generation of cadmium sulfide | |
| CN110346438B (en) | Based on PbS/Co3O4Preparation method of composite signal attenuation type photoelectrochemical immunosensor | |
| CN110261448A (en) | A kind of preparation method and application of the signal suppressive optical electro-chemistry Procalcitonin sensor based on zinc titanium composite material | |
| CN110308187B (en) | Preparation method and application of photoelectrochemical aptamer sensor based on zinc and cobalt doped porous nano bismuth vanadate/bismuth sulfide | |
| CN110118759B (en) | Oxytetracycline fluorescence detection method based on surface passivation and covalent coupling | |
| CN110487868B (en) | Method for detecting chlorpyrifos based on photoelectrochemical sensor | |
| CN113203788A (en) | Bimodal rapid detection method for acetamiprid in fruits and vegetables | |
| CN113588752B (en) | Preparation method and application of electrochemiluminescence aptamer sensor | |
| CN110297023B (en) | Preparation method and application of self-enhanced photoelectrochemical immunosensor for detecting procalcitonin electrochemical catalysis assistance | |
| CN107121462A (en) | A kind of preparation method for vulcanizing the dual decrease cadmium sulfide of Cu/SiO 2/carbon doping titanium dioxide insulin optical electro-chemistry sensor | |
| CN106198501B (en) | A kind of preparation method for the Electrochemiluminescsensor sensor detecting aflatoxin | |
| Dong et al. | Research progress on photoelectrochemical sensors for contamination analysis in agricultural fields | |
| Sun | A potentiometric immunosensor for enterovirus 71 based on bis-MPA-COOH dendrimer-doped AgCl nanospheres with a silver ion-selective electrode |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20190820 Termination date: 20200803 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |