CN105717099B - A kind of preparation method and application of electrogenerated chemiluminescence furazolidone biology sensor - Google Patents
A kind of preparation method and application of electrogenerated chemiluminescence furazolidone biology sensor Download PDFInfo
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Abstract
The invention discloses a kind of preparation method of electrogenerated chemiluminescence furazolidone biology 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 nano composite material Mn TiO2/g‑C3N4, good biocompatibility and big specific surface area using the material, furazolidone antibody in load, then the crosslinked action horseradish peroxidase of glutaraldehyde is passed through, when being detected, because horseradish peroxidase can produce oxygen so that catalyzing hydrogen peroxide is in situ, and and then to be electrochemically reacted with bottom liquid, produce high and stable electrochemiluminescence signal, antibody is recycled to be combined the influence to electron transport ability with the specific quantification of antigen, so that electrochemiluminescence signal accordingly reduces, it is low that cost finally has been made, high sensitivity, specificity is good, detection is quick, prepare the electroluminescent chemiluminescence biosensor of simple detection furazolidone.
Description
Technical field
The present invention relates to a kind of preparation method of electrogenerated chemiluminescence furazolidone biology sensor.Belong to novel nano work(
Can material and biosensor technology field.
Background technology
Furazolidone（Furazolidone）It is a kind of Nitrofuran antibiotics, is broad spectrum antibiotic.As veterinary drug in use, furan
Oxazolone of muttering has good drug effect to preventing and treating some protozoosises, saprolegniasis, bacterial gill rot disease, red skin disease, hemorrhage etc..In aquaculture
In, furazolidone can be used for treatment animal and bird intestines infection, as piglet is yellow, dysentery characterized by white mucous stool.In aquatic products industry, furazolidone is to Salmonoidei sense
Dye Myxosoma cerebralis has certain curative effect.But furazolidone is classified as the medicine prohibitted the use of by the Ministry of Agriculture of China, must not be eaten in animality
Detected in product.FDA also prohibited itrofurans in 2002（Including furazolidone）Use in animal food.
At present, the method for detecting furazolidone mainly has chromatography, mass spectrography etc..Such method instrument is valuable, operation is multiple
Miscellaneous, laboratory personnel could be detected after needing professional training.Therefore, R＆D costs are low, detection is fast, high sensitivity, specificity
Strong furazolidone sensor is significant.
Electrochemiluminescsensor sensor due to its high sensitivity, specificity it is good, easy to operate the advantages that be widely used in
The fields such as clinical diagnosis, Pharmaceutical Analysis, environmental monitoring.The superior Electrochemiluminescsensor sensor of processability, its most critical skill
Art is exactly the raising of the performance such as luminous intensity and stability and effective fixation of immune molecule and reappearance.Titanium dioxide is application
A kind of most commonly used photocatalyst material, because sheet-like titanium dioxide nanomaterial can expose more high miller index surfaces,
With higher photocatalytic activity, titanium dioxide nanoplate has than nano-particle preferably application prospect, for titanium dioxide
The research of nanometer sheet also receives much concern.Meanwhile titanium dioxide poorly conductive also limit by single titanium dioxide nano material structure
The sensitivity for the Electrochemiluminescsensor sensor built is not universal high, is unfavorable for practical application.But on semiconductor nano material
Modification or compound special nano material, the Resonance energy transfer of semiconductor surface can be effectively improved, produce stronger light
Intensity, and greatly improve detection sensitivity.Therefore, designing, preparing efficient, stable titanium dioxide nanoplate and its trim is
Prepare the key technology of Electrochemiluminescsensor sensor.
The content of the invention
Prepare that simple, high sensitivity, detection be quick, electroluminescent chemistry of high specificity it is an object of the invention to provide a kind of
The preparation method of luminous furazolidone biology sensor, prepared sensor, quick, the sensitive inspection available for furazolidone
Survey.Based on this purpose, the method comprises the steps of firstly, preparing a kind of New Two Dimensional nano composite material, i.e. additive Mn titanium dioxide nanoplate
In-situ reaction carbonitride two-dimensional nano composite Mn-TiO2/g-C3N4, utilize the good biocompatibility of the material and big
Specific surface area, furazolidone antibody in load, horseradish peroxidase is then fixed by the crosslinked action of glutaraldehyde, entered
During row detection, because horseradish peroxidase can be with catalyzing hydrogen peroxide in-situ preparation O2, and with the K in the liquid of bottom2S2O8Carry out electricity
Chemical reaction, electrochemiluminescence signal is produced, recycle antibody to be combined with the specific quantification of antigen to electron transport ability
Influence so that current strength reduces, and so as to reduce luminous intensity, finally realizes using unmarked electrogenerated chemiluminescence side
The structure of the biology sensor of method detection furazolidone.
The technical solution adopted by the present invention is as follows：
1. a kind of preparation method of electrogenerated chemiluminescence furazolidone biology sensor, described electrogenerated chemiluminescence furan
Oxazolone biology sensor mutter by working electrode, Mn-TiO2/g-C3N4, it is furazolidone antibody, bovine serum albumin(BSA), glutaraldehyde, peppery
Root peroxidase forms；Described Mn-TiO2/g-C3N4For additive Mn titanium dioxide nanoplate In-situ reaction carbonitride two dimension
Nano composite material；
Characterized in that, described preparation method includes following preparation process：
a. Mn-TiO2/g-C3N4Preparation；
B. the preparation of electrogenerated chemiluminescence furazolidone biology sensor；
Wherein, step a prepares Mn-TiO2/g-C3N4Concretely comprise the following steps：
First, 0.8 ~ 1.2 mmol manganese salts are taken to be added in 5 mL butyl titanates, in whipping process, it is slowly added to 0.5 ~
0.8 mL hydrofluoric acid, react in a kettle 18 ~ 24 hours, after being cooled to room temperature at 160 ~ 200 DEG C, with ultra-pure water and anhydrous
Ethanol centrifuge washing three times after, be dried in vacuo at 50 DEG C；Secondly, the dried solids of 150 ~ 250 mg and 400mg melamines are taken
Amine mixes, and grind into powder；Then, the powder of grinding is put into Muffle furnace, programming rate is 1 ~ 3 DEG C/min, 480
Calcined 0.5 ~ 5 hour at ~ 560 DEG C；Finally, the powder after calcining is cooled to room temperature, that is, Mn-TiO is made2/g-C3N4；
Described manganese salt is selected from one of following：Manganese sulfate, manganese chloride, manganese nitrate；
Step b prepares concretely comprising the following steps for electrogenerated chemiluminescence furazolidone biology sensor：
（1）Using ITO electro-conductive glass as working electrode, in the μ L of electrode surface drop coating 8 ~ 12 Mn-TiO2/g-C3N4Colloidal sol,
Dry at room temperature；
（2）By step（1）In obtained electrode cushioning liquid PBS, continue in the μ L of electrode surface drop coating 8 ~ 12
10 μ g/mL furazolidone antibody-solutions, preserve in 4 DEG C of refrigerators and dry；
（3）By step（2）In obtained electrode PBS, it is 100 to continue in the μ L concentration of electrode surface drop coating 8 ~ 12
μ g/mL bovine serum albumin solution, preserve in 4 DEG C of refrigerators and dry；
（4）By step（3）In obtained electrode PBS, the glutaraldehyde continued in the μ L of electrode surface drop coating 2 ~ 4 is molten
Liquid, preserve in 4 DEG C of refrigerators and dry；
（5）By step（4）In obtained electrode PBS, it is 20 μ to continue in the μ L concentration of electrode surface drop coating 6 ~ 10
G/mL horseradish peroxidase solution, preserve in 4 DEG C of refrigerators and dry；
（6）By step（5）In obtained electrode PBS, preserved in 4 DEG C of refrigerators after drying, that is, be made electroluminescent
Chemiluminescence furazolidone biology sensor；
Described Mn-TiO2/g-C3N4Sol solutionses are by 50 mg Mn-TiO2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters
In, and the obtained hydrosol after 30 min of ultrasound；
Described PBS is 10mmol/L phosphate buffer solution, and the pH value of described phosphate buffer solution is 7.4；
Described glutaraldehyde solution is the glutaraldehyde water solution that volume ratio is 2.5%.
2. the application of the electrogenerated chemiluminescence furazolidone biology sensor prepared by preparation method of the present invention,
It is characterised in that it includes following applying step：
A. standard liquid is prepared：Prepare the furazolidone standard liquid of one group of various concentrations including blank standard specimen；
B. working electrode is modified：The electrogenerated chemiluminescence furazolidone bio-sensing prepared by method will be prepared as above
The furazolidone standard liquid for the various concentrations prepared in step a is distinguished drop coating to working electrode table by device as working electrode
Face, preserve in 4 DEG C of refrigerators；
C. working curve is drawn：Using Ag/AgCl electrodes as reference electrode, platinum electrode is used as to electrode, with step b
The working electrode composition three-electrode system modified, is connected in electrochemiluminescdetection detection equipment；In a cell successively
Add 15 mL K2S2O8The H of solution and 100 μ L2O2Solution；Circulation electricity is applied to the working electrode of assembling with cyclic voltammetry
Pressure；According to the relation between the light signal strength of the electrogenerated chemiluminescence of gained and furazolidone antigen concentration of standard solution, paint
Working curve processed；The light signal strength of blank standard specimen is designated asD 0, the optical signal of the furazolidone standard liquid containing various concentrations
Intensity is designated asD i, the difference that response light signal strength reduces is ΔD = D 0-D i, ΔDIt is dense with the quality of furazolidone standard liquid
DegreeCBetween it is linear, draw ΔD-CWorking curve；
D. the detection of furazolidone：The furazolidone standard liquid in step a is replaced with testing sample, according to step b and
Method in c is detected, the difference DELTA reduced according to response light signal strengthDAnd working curve, obtain furan in testing sample
Mutter the content of oxazolone；
Described K2S2O8Solution is by 1 mol K2S2O8It is dissolved in the cushioning liquid of 10 L pH=6.5 and makes with 1 mol KCl
Standby to obtain, the cushioning liquid of described pH=6.5 is the phosphate buffer solution that pH value is 6.5；
Described H2O2Solution is aqueous hydrogen peroxide solution, and the concentration of described aqueous hydrogen peroxide solution is 10%.
The useful achievement of the present invention
（1）Electrogenerated chemiluminescence furazolidone biology sensor of the present invention is prepared simply, easy to operate, is realized
To the quick, sensitive of sample, high selectivity detection, and cost is low, can be applied to portable inspectiont, before having market development
Scape；
（2）The present invention is prepared for New-type matrix material Mn-TiO first2/g-C3N4, because manganese is on titanium dioxide nanoplate
Growth in situ and fully contacted with titanium dioxide nanoplate, using manganese metal surface plasma body act on, effectively increase
Semiconductor substrate Resonance energy transfer ability, although solving, titanium dioxide nanoplate specific surface area is bigger and mesoporous characteristic is fitted
For electrogenerated chemiluminescence host material, but the technical problem that electrochemiluminescence signal is unstable；Simultaneously because carbonitride
g-C3N4Good electric conductivity and electrogenerated chemiluminescence characteristic, along with titanium dioxide nanoplate is fully dispersed thereon,
Electrogenerated chemiluminescence intensity is greatly increased, titanium dioxide nanoplate poorly conductive is solved and electrogenerated chemiluminescence intensity is weak
And it is unfavorable for preparing the technical problem of Electrochemiluminescsensor sensor, therefore, effective preparation of the material, there is important science
Meaning and application value；
（3）The present invention is first by Mn-TiO2/g-C3N4Applied in the preparation of electroluminescent chemiluminescence biosensor, significantly
The strength and stability of electrogenerated chemiluminescence is improved, the detection sensitivity of Electrochemiluminescsensor sensor is substantially increased, makes
Obtaining electroluminescent chemiluminescence biosensor realizes the application in real work；The application of the material, also passed for associated biomolecule
Sensor, such as optical electro-chemistry sensor, electrochemical sensor provide Technical Reference, have extensive potential use value.
Embodiment
The Mn-TiO of embodiment 12/g-C3N4Preparation
First, take 0.8 mmol manganese salts to be added in 5 mL butyl titanates, in whipping process, be slowly added to 0.5 mL hydrogen
Fluoric acid, react in a kettle 24 hours, after being cooled to room temperature at 160 DEG C, with ultra-pure water and absolute ethyl alcohol centrifuge washing three times
Afterwards, it is dried in vacuo at 50 DEG C；Secondly, the dried solids of 150 mg are taken to be mixed with 400 mg melamines, and pulverize
End；Then, the powder of grinding is put into Muffle furnace, programming rate is 1 DEG C/min, is calcined 5 hours at 480 DEG C；Finally,
Powder after calcining is cooled to room temperature, that is, Mn-TiO is made2/g-C3N4；
Described manganese salt is manganese sulfate.
The Mn-TiO of embodiment 22/g-C3N4Preparation
First, take 1.0 mmol manganese salts to be added in 5 mL butyl titanates, in whipping process, be slowly added to 0.6 mL hydrogen
Fluoric acid, react in a kettle 21 hours, after being cooled to room temperature at 180 DEG C, with ultra-pure water and absolute ethyl alcohol centrifuge washing three times
Afterwards, it is dried in vacuo at 50 DEG C；Secondly, the dried solids of 200 mg are taken to be mixed with 400 mg melamines, and pulverize
End；Then, the powder of grinding is put into Muffle furnace, programming rate is 2 DEG C/min, is calcined 2 hours at 520 DEG C；Finally,
Powder after calcining is cooled to room temperature, that is, Mn-TiO is made2/g-C3N4；
Described manganese salt is manganese chloride.
The Mn-TiO of embodiment 32/g-C3N4Preparation
First, take 1.2 mmol manganese salts to be added in 5 mL butyl titanates, in whipping process, be slowly added to 0.8 mL hydrogen
Fluoric acid, react in a kettle 18 hours, after being cooled to room temperature at 200 DEG C, with ultra-pure water and absolute ethyl alcohol centrifuge washing three
After secondary, it is dried in vacuo at 50 DEG C；Secondly, the dried solids of 250 mg are taken to be mixed with 400 mg melamines, and pulverize
End；Then, the powder of grinding is put into Muffle furnace, programming rate is 3 DEG C/min, is calcined 0.5 hour at 560 DEG C；Most
Afterwards, the powder after calcining is cooled to room temperature, that is, Mn-TiO is made2/g-C3N4；
Described manganese salt is manganese nitrate.
The preparation method of the electrogenerated chemiluminescence furazolidone biology sensor of embodiment 4
（1）Using a width of 1 cm, a length of 4 cm ITO electro-conductive glass as working electrode, the μ L's of electrode surface drop coating 8
Mn-TiO2/g-C3N4Colloidal sol, dry at room temperature；
（2）By step（1）In obtained electrode cushioning liquid PBS, continue in the μ of 8 μ L of electrode surface drop coating 10
G/mL furazolidone antibody-solutions, preserve in 4 DEG C of refrigerators and dry；
（3）By step（2）In obtained electrode PBS, it is 100 μ to continue in the μ L concentration of electrode surface drop coating 8
G/mL bovine serum albumin solution, preserve in 4 DEG C of refrigerators and dry；
（4）By step（3）In obtained electrode PBS, the glutaraldehyde continued in the μ L of electrode surface drop coating 2 is molten
Liquid, preserve in 4 DEG C of refrigerators and dry；
（5）By step（4）In obtained electrode PBS, it is 20 μ g/ to continue in the μ L concentration of electrode surface drop coating 6
ML horseradish peroxidase solution, preserve in 4 DEG C of refrigerators and dry；
（6）By step（5）In obtained electrode PBS, preserved in 4 DEG C of refrigerators after drying, that is, be made electroluminescent
Chemiluminescence furazolidone biology sensor；
Described Mn-TiO2/g-C3N4Colloidal sol is by 50 mg Mn-TiO2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters
In, and the obtained hydrosol after 30 min of ultrasound；
Described PBS is 10mmol/L phosphate buffer solution, and the pH value of described phosphate buffer solution is 7.4；
Described glutaraldehyde solution is the glutaraldehyde water solution that volume ratio is 2.5%.
The preparation method of the electrogenerated chemiluminescence furazolidone biology sensor of embodiment 5
（1）Using a width of 1 cm, a length of 4 cm ITO electro-conductive glass as working electrode, the μ L's of electrode surface drop coating 10
Mn-TiO2/g-C3N4Colloidal sol, dry at room temperature；
（2）By step（1）In obtained electrode cushioning liquid PBS, continue in the μ L 10 of electrode surface drop coating 10
μ g/mL furazolidone antibody-solutions, preserve in 4 DEG C of refrigerators and dry；
（3）By step（2）In obtained electrode PBS, it is 100 μ to continue in the μ L concentration of electrode surface drop coating 10
G/mL bovine serum albumin solution, preserve in 4 DEG C of refrigerators and dry；
（4）By step（3）In obtained electrode PBS, the glutaraldehyde continued in the μ L of electrode surface drop coating 3 is molten
Liquid, preserve in 4 DEG C of refrigerators and dry；
（5）By step（4）In obtained electrode PBS, it is 20 μ g/ to continue in the μ L concentration of electrode surface drop coating 8
ML horseradish peroxidase solution, preserve in 4 DEG C of refrigerators and dry；
（6）By step（5）In obtained electrode PBS, preserved in 4 DEG C of refrigerators after drying, that is, be made electroluminescent
Chemiluminescence furazolidone biology sensor；
Described Mn-TiO2/g-C3N4Colloidal sol is by 50 mg Mn-TiO2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters
In, and the obtained hydrosol after 30 min of ultrasound；
Described PBS is 10 mmol/L phosphate buffer solution, and the pH value of described phosphate buffer solution is 7.4；
Described glutaraldehyde solution is the glutaraldehyde water solution that volume ratio is 2.5%.
The preparation method of the electrogenerated chemiluminescence furazolidone biology sensor of embodiment 6
（1）Using a width of 1 cm, a length of 4 cm ITO electro-conductive glass as working electrode, the μ L's of electrode surface drop coating 12
Mn-TiO2/g-C3N4Colloidal sol, dry at room temperature；
（2）By step（1）In obtained electrode cushioning liquid PBS, continue in the μ L 10 of electrode surface drop coating 12
μ g/mL furazolidone antibody-solutions, preserve in 4 DEG C of refrigerators and dry；
（3）By step（2）In obtained electrode PBS, it is 100 μ to continue in the μ L concentration of electrode surface drop coating 12
G/mL bovine serum albumin solution, preserve in 4 DEG C of refrigerators and dry；
（4）By step（3）In obtained electrode PBS, the glutaraldehyde continued in the μ L of electrode surface drop coating 4 is molten
Liquid, preserve in 4 DEG C of refrigerators and dry；
（5）By step（4）In obtained electrode PBS, it is 20 μ g/ to continue in the μ L concentration of electrode surface drop coating 10
ML horseradish peroxidase solution, preserve in 4 DEG C of refrigerators and dry；
（6）By step（5）In obtained electrode PBS, preserved in 4 DEG C of refrigerators after drying, that is, be made electroluminescent
Chemiluminescence furazolidone biology sensor；
Described Mn-TiO2/g-C3N4Colloidal sol is by 50 mg Mn-TiO2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters
In, and the obtained hydrosol after 30 min of ultrasound；
Described PBS is 10 mmol/L phosphate buffer solution, and the pH value of described phosphate buffer solution is 7.4；
Described glutaraldehyde solution is the glutaraldehyde water solution that volume ratio is 2.5%.
Electrogenerated chemiluminescence furazolidone biology sensor prepared by the embodiment 1 ~ 6 of embodiment 7, applied to furazolidone
Detection, step is as follows：
（1）Standard liquid is prepared：Prepare the furazolidone standard liquid of one group of various concentrations including blank standard specimen；
（2）Working electrode is modified：The electrogenerated chemiluminescence furazolidone bio-sensing prepared by method will be prepared as above
Device is as working electrode, by step（1）The furazolidone standard liquid of the various concentrations of middle preparation distinguishes drop coating to working electrode
Surface, preserve 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（2）The working electrode composition three-electrode system modified, is connected in electrochemiluminescdetection detection equipment；In a cell
Successively add 15 mL K2S2O8The H of solution and 100 μ L2O2Solution；The working electrode of assembling is applied with cyclic voltammetry and followed
Loop voltag；According to the pass between the light signal strength of the electrogenerated chemiluminescence of gained and furazolidone antigen concentration of standard solution
System, drawing curve；The light signal strength of blank standard specimen is designated asD 0, the light of the furazolidone standard liquid containing various concentrations
Signal intensity is designated asD i, the difference that response light signal strength reduces is ΔD = D 0-D i, ΔDWith the matter of furazolidone standard liquid
Measure concentrationCBetween it is linear, draw ΔD-CWorking curve；The linear detection range of furazolidone is：0.003~100
Ng/mL, detection are limited to：1.1 pg/mL；
（4）The detection of furazolidone：The furazolidone standard liquid in step a is replaced with testing sample, according to step b and
Method in c is detected, the difference DELTA reduced according to response light signal strengthDAnd working curve, obtain furan in testing sample
Mutter the content of oxazolone；
Described K2S2O8Solution is by 1 mol K2S2O8It is dissolved in the cushioning liquid of 10 L pH=6.5 and makes with 1 mol KCl
Standby to obtain, the cushioning liquid of described pH=6.5 is the phosphate buffer solution that pH value is 6.5；
Described H2O2Solution is aqueous hydrogen peroxide solution, and the concentration of described aqueous hydrogen peroxide solution is 10%.
Claims (2)
1. a kind of preparation method of electrogenerated chemiluminescence furazolidone biology sensor, described electrogenerated chemiluminescence furazolidone
Biology sensor is by working electrode, Mn-TiO2/g-C3N4, furazolidone antibody, bovine serum albumin(BSA), glutaraldehyde, horseradish peroxide
Compound enzyme forms；Described Mn-TiO2/g-C3N4Answered for additive Mn titanium dioxide nanoplate In-situ reaction carbonitride two-dimensional nano
Condensation material；
Characterized in that, described preparation method includes following preparation process：
a. Mn-TiO2/g-C3N4Preparation；
B. the preparation of electrogenerated chemiluminescence furazolidone biology sensor；
Wherein, step a prepares Mn-TiO2/g-C3N4Concretely comprise the following steps：
First, take 0.8 ~ 1.2 mmol manganese salts to be added in 5 mL butyl titanates, in whipping process, be slowly added to 0.5 ~ 0.8
ML hydrofluoric acid, react in a kettle 18 ~ 24 hours, after being cooled to room temperature at 160 ~ 200 DEG C, with ultra-pure water and absolute ethyl alcohol
Centrifuge washing three times after, be dried in vacuo at 50 DEG C；Secondly, the dried solids of 150 ~ 250 mg are taken to be mixed with 400mg melamines
Close, and grind into powder；Then, the powder of grinding is put into Muffle furnace, programming rate is 1 ~ 3 DEG C/min, 480 ~ 560
Calcined 0.5 ~ 5 hour at DEG C；Finally, the powder after calcining is cooled to room temperature, that is, Mn-TiO is made2/g-C3N4；
Described manganese salt is selected from one of following：Manganese sulfate, manganese chloride, manganese nitrate；
Step b prepares concretely comprising the following steps for electrogenerated chemiluminescence furazolidone biology sensor：
（1）Using ITO electro-conductive glass as working electrode, in the μ L of electrode surface drop coating 8 ~ 12 Mn-TiO2/g-C3N4Colloidal sol, room temperature
Under dry；
（2）By step（1）In obtained electrode cushioning liquid PBS, continue in the μ of 8 ~ 12 μ L of electrode surface drop coating 10
G/mL furazolidone antibody-solutions, preserve in 4 DEG C of refrigerators and dry；
（3）By step（2）In obtained electrode PBS, it is 100 μ g/ to continue in the μ L concentration of electrode surface drop coating 8 ~ 12
ML bovine serum albumin solution, preserve in 4 DEG C of refrigerators and dry；
（4）By step（3）In obtained electrode PBS, continue the glutaraldehyde solution in the μ L of electrode surface drop coating 2 ~ 4,4
Preserve and dry in DEG C refrigerator；
（5）By step（4）In obtained electrode PBS, it is 20 μ g/mL to continue in the μ L concentration of electrode surface drop coating 6 ~ 10
Horseradish peroxidase solution, preserve in 4 DEG C of refrigerators and dry；
（6）By step（5）In obtained electrode PBS, preserved in 4 DEG C of refrigerators after drying, that is, electroluminescent chemistry be made
Luminous furazolidone biology sensor；
Described Mn-TiO2/g-C3N4Colloidal sol is by 50 mg Mn-TiO2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters, and
The obtained hydrosol after 30 min of ultrasound；
Described PBS is 10mmol/L phosphate buffer solution, and the pH value of described phosphate buffer solution is 7.4；
Described glutaraldehyde solution is the glutaraldehyde water solution that volume ratio is 2.5%.
2. the application of the electrogenerated chemiluminescence furazolidone biology sensor prepared by preparation method as claimed in claim 1,
It is characterised in that it includes following applying step：
A. standard liquid is prepared：Prepare the furazolidone standard liquid of one group of various concentrations including blank standard specimen；
B. working electrode is modified：Electrogenerated chemiluminescence furazolidone prepared by preparation method as claimed in claim 1 is given birth to
The furazolidone standard liquid for the various concentrations prepared in step a is distinguished drop coating to work by thing sensor as working electrode
Electrode surface, preserve in 4 DEG C of refrigerators；
C. working curve is drawn：It is used as Ag/AgCl electrode as reference electrode, platinum electrode to electrode, is repaiied with step b
The working electrode composition three-electrode system adornd, is connected in electrochemiluminescdetection detection equipment；Successively add in a cell
15 mL K2S2O8The H of solution and 100 μ L2O2Solution；Cyclical voltage is applied to the working electrode of assembling with cyclic voltammetry；
According to the relation between the light signal strength of the electrogenerated chemiluminescence of gained and furazolidone antigen concentration of standard solution, work is drawn
Make curve；The light signal strength of blank standard specimen is designated asD 0, the light signal strength of the furazolidone standard liquid containing various concentrations
It is designated asD i, the difference that response light signal strength reduces is ΔD = D 0-D i, ΔDWith the mass concentration of furazolidone standard liquidC
Between it is linear, draw ΔD-CWorking curve；
D. the detection of furazolidone：The furazolidone standard liquid in step a is replaced with testing sample, according in step b and c
Method detected, according to response light signal strength reduce difference DELTADAnd working curve, obtain furans azoles in testing sample
The content of ketone；
Described K2S2O8Solution is by 1 mol K2S2O8It is dissolved in the cushioning liquid of 10 L pH=6.5 and is prepared into 1 mol KCl
Arrive, the cushioning liquid of described pH=6.5 is the phosphate buffer solution that pH value is 6.5；
Described H2O2Solution is aqueous hydrogen peroxide solution, and the concentration of described aqueous hydrogen peroxide solution is 10%.
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| CN101464462A (en) * | 2008-10-24 | 2009-06-24 | 山东大学 | Chemical luminescence ELISA detection reagent kit for furazolidone |
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| CN104374910A (en) * | 2014-11-19 | 2015-02-25 | 无锡中德伯尔生物技术有限公司 | Kit and method applied to detection of nitrofurans drug metabolite |
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| US3980434A (en) * | 1975-07-30 | 1976-09-14 | Morton-Norwich Products, Inc. | Method for determining furazolidone in animal tissue |
| CN101464462A (en) * | 2008-10-24 | 2009-06-24 | 山东大学 | Chemical luminescence ELISA detection reagent kit for furazolidone |
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| CN104374910A (en) * | 2014-11-19 | 2015-02-25 | 无锡中德伯尔生物技术有限公司 | Kit and method applied to detection of nitrofurans drug metabolite |
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