CN105717099A - Preparation method and application of electrogenerated chemiluminescence furazolidone biosensor - Google Patents
Preparation method and application of electrogenerated chemiluminescence furazolidone biosensor Download PDFInfo
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Abstract
The invention discloses a preparation method of an electrogenerated chemiluminescence furazolidone biosensor and belongs to the technical field of novel nano functional materials and biological sensors. A novel two-dimensional nano composite material Mn-TiO2/g-C3N4 is prepared first, the material is loaded with furazolidone antibodies by means of good biocompatibility and large specific surface area, then by means of the cross-linking effect of glutaraldehyde, horse radish peroxidase can catalyze hydrogen peroxide to generate oxygen enzyme in situ in detection, an electrochemical reaction is performed on a base solution, a high and stable electrogenerated chemiluminescence signal is generated, by means of influences of specific quantitative combination of antibodies and antigens on electron transmission capacity, the electrogenerated chemiluminescence signal is correspondingly reduced, finally the low-cost, high-sensitivity and good-specificity electrogenerated chemiluminescence biosensor for detecting furazolidone is prepared, and the biosensor is fast in detection and easy to prepare.
Description
Technical field
The present invention relates to the preparation method of a kind of electrogenerated chemiluminescence furazolidone biosensor.Belong to Nano-function thin films and biosensor technology field.
Background technology
Furazolidone (furazolidone) is a kind of Nitrofuran antibiotics, for broad spectrum antibiotic.When using as veterinary drug, furazolidone has good drug effect to preventing and treating some protozoacide, fish molds, bacterial gill rot disease, erythroderma, hemorrhagic disease etc..In aquaculture, furazolidone can be used for treating animal and bird intestines to be infected, as yellow in piglet, Hakuri.In aquatic products industry, Salmonoidei is infected Myxosoma cerebralis by furazolidone certain curative effect.But furazolidone is classified as the medicine prohibitted the use of by the Ministry of Agriculture of China, must not detect in animal food.FDA also prohibited the itrofurans (including furazolidone) use in animal food in 2002.
At present, the method for detection furazolidone mainly has chromatography, mass spectrography etc..This type of method instrument is valuable, operation complexity, and laboratory personnel just can detect after needing professional training.Therefore, R&D costs are low, it is fast, highly sensitive to detect, the furazolidone sensor of high specificity is significant.
Electrochemiluminescsensor sensor is widely used in the fields such as clinical diagnosis, pharmaceutical analysis, environmental monitoring due to advantages such as it are highly sensitive, specificity is good, easy and simple to handle.The Electrochemiluminescsensor sensor that processability is superior, its most critical technology is exactly the raising of the performances such as the most fixing and repeatability of luminous intensity and stability and immune molecule.Titanium dioxide is a kind of photocatalyst material being most widely used, owing to sheet-like titanium dioxide nanomaterial can expose more high miller index surface, there is higher photocatalytic activity, titanium dioxide nanoplate has ratio nanoparticle preferably application prospect, and the research for titanium dioxide nanoplate also receives much concern.Meanwhile, the sensitivity that titanium dioxide poorly conductive also limit the Electrochemiluminescsensor sensor built by single titanium dioxide nano material is the highest, is unfavorable for actual application.But, semiconductor nano material modified or be combined special nano material, the Resonance energy transfer of semiconductor surface can be effectively improved, producing higher luminous intensity, and be greatly improved detection sensitivity.Therefore, design, prepare efficient, stable titanium dioxide nanoplate and trim is the key technology preparing Electrochemiluminescsensor sensor.
Summary of the invention
It is an object of the invention to provide a kind of prepare simple, highly sensitive, detection quickly, the preparation method of the electrogenerated chemiluminescence furazolidone biosensor of high specificity, prepared sensor, can be used for quick, the Sensitive Detection of furazolidone.Based on this purpose, 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-TiO are the method comprises the steps of firstly, preparing2/g-C3N4Utilize the good biocompatibility of this material and big specific surface area, furazolidone antibody in load, then fix horseradish peroxidase by the crosslinked action of glutaraldehyde, when detecting, owing to horseradish peroxidase can be with catalyzing hydrogen peroxide in-situ preparation O2, and with the K in end liquid2S2O8It is electrochemically reacted, produce electrochemiluminescence signal, recycling antibody is combined the impact on electron transport ability with the specific quantification of antigen, current intensity is reduced, thus reduce luminous intensity, finally achieve the structure of the biosensor using unmarked electrogenerated chemiluminescence method detection furazolidone.
The technical solution used in the present invention is as follows:
1. a preparation method for electrogenerated chemiluminescence furazolidone biosensor, described electrogenerated chemiluminescence furazolidone biosensor is by working electrode, Mn-TiO2/g-C3N4, furazolidone antibody, bovine serum albumin, glutaraldehyde, horseradish peroxidase composition；Described Mn-TiO2/g-C3N4For additive Mn titanium dioxide nanoplate In-situ reaction carbonitride two-dimensional nano composite；
It is characterized in that, described preparation method includes following preparation process:
a. Mn-TiO2/g-C3N4Preparation；
B. the preparation of electrogenerated chemiluminescence furazolidone biosensor；
Wherein, step a prepares Mn-TiO2/g-C3N4Concretely comprise the following steps:
First, take 0.8 ~ 1.2 mmol manganese salt and join in 5 mL butyl titanates, in whipping process, be slowly added to 0.5 ~ 0.8
ML Fluohydric acid., reacts at 160 ~ 200 DEG C 18 ~ 24 hours, after being cooled to room temperature in a kettle., after ultra-pure water and dehydrated alcohol centrifuge washing three times, is vacuum dried at 50 DEG C；Secondly, 150 ~ 250 are taken
The dried solid of mg mixes with 400mg tripolycyanamide, and grind into powder；Then, putting in Muffle furnace by the powder of grinding, programming rate is 1 ~ 3 DEG C/min, calcines 0.5 ~ 5 hour at 480 ~ 560 DEG C；Finally, the powder after calcining is cooled to room temperature, i.e. prepares Mn-TiO2/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 of electrogenerated chemiluminescence furazolidone biosensor:
(1) with ITO electro-conductive glass as working electrode, at the Mn-TiO of electrode surface drop coating 8 ~ 12 L2/g-C3N4Colloidal sol, dries under room temperature；
(2) the electrode buffer solution PBS that will obtain in step (1), continues the furazolidone antibody-solutions at electrode surface drop coating 8 ~ 12 L 10 g/mL, preserves and dry in 4 DEG C of refrigerators；
(3) the electrode PBS that will obtain in step (2), continues at the bovine serum albumin solution that electrode surface drop coating 8 ~ 12 L concentration is 100 g/mL, preserves and dry in 4 DEG C of refrigerators；
(4) the electrode PBS that will obtain in step (3), continues the glutaraldehyde solution at electrode surface drop coating 2 ~ 4 L, preserves and dry in 4 DEG C of refrigerators；
(5) the electrode PBS that will obtain in step (4), continues at the horseradish peroxidase solution that electrode surface drop coating 6 ~ 10 L concentration is 20 g/mL, preserves and dry in 4 DEG C of refrigerators；
(6) the electrode PBS that will obtain in step (5), preserves in 4 DEG C of refrigerators after drying, i.e. prepares electrogenerated chemiluminescence furazolidone biosensor；
Described Mn-TiO2/g-C3N4Solution is by the Mn-TiO of 50 mg2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters, and the hydrosol prepared after ultrasonic 30 min；
Described PBS is the phosphate buffered solution of 10mmol/L, and the pH value of described phosphate buffered solution is 7.4；
Described glutaraldehyde solution be volume ratio be the glutaraldehyde water solution of 2.5%.
The application of the electrogenerated chemiluminescence furazolidone biosensor prepared by preparation method the most of the present invention, it is characterised in that include following applying step:
A. standard solution preparation: prepare the furazolidone standard solution of one group of variable concentrations including blank standard specimen；
B. working electrode is modified: be working electrode by the electrogenerated chemiluminescence furazolidone biosensor prepared by preparation method as claimed in claim 1, by the drop coating respectively of the furazolidone standard solution of the variable concentrations of preparation in step a to working electrode surface, 4 DEG C of refrigerators preserve；
C. working curve is drawn:, be connected on electrochemiluminescdetection detection equipment as reference electrode, platinum electrode as to electrode, the working electrode composition three-electrode system modified with step b by Ag/AgCl electrode；Successively add the K of 15 mL in a cell2S2O8Solution and the H of 100 L2O2Solution；Cyclical voltage is applied with the cyclic voltammetry working electrode to assembling；Relation between light signal strength and the furazolidone antigen concentration of standard solution of the electrogenerated chemiluminescence according to gained, drawing curve；The light signal strength of blank standard specimen is designated asD 0, the light signal strength of the furazolidone standard solution containing variable concentrations is designated asD i, the difference that response light signal intensity reduces is ΔD =D 0-D i, ΔDMass concentration with furazolidone standard solutionCBetween linear, draw ΔD-CWorking curve；
D. the detection of furazolidone: replace the furazolidone standard solution in step a with testing sample, detect according to the method in step b and c, the difference DELTA reduced according to response light signal intensityDAnd working curve, obtain the content of furazolidone in testing sample；
Described K2S2O8Solution is by 1 mol K2S2O8Be dissolved in the pH=6.5 buffer solution of 10 L with 1 mol KCl and preparing, described pH=6.5 buffer solution be pH value be the phosphate buffered solution of 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 biosensor of the present invention preparation is simple, easy to operate, it is achieved that the selective enumeration method quick, sensitive, high to sample, and low cost, can be applicable to portable inspectiont, has market development prospect；
(2) present invention is prepared for New-type matrix material Mn-TiO first2/g-C3N4Fully contact with titanium dioxide nanoplate due to manganese growth in situ on titanium dioxide nanoplate, utilize the metal surface plasma body effect of manganese, it is effectively increased semiconductor substrate Resonance energy transfer ability, solve that although titanium dioxide nanoplate specific surface area is bigger and mesoporous characteristic is applicable to electrogenerated chemiluminescence host material, but the technical problem that electrochemiluminescence signal is unstable；Simultaneously because carbonitride g-C3N4Good electric conductivity and the characteristic of electrogenerated chemiluminescence, add titanium dioxide nanoplate thereon fully dispersed, greatly increase electrogenerated chemiluminescence intensity, solve titanium dioxide nanoplate poorly conductive and electrogenerated chemiluminescence intensity is weak and be unfavorable for preparing the technical problem of Electrochemiluminescsensor sensor, therefore, effective preparation of this material, has important scientific meaning and using value；
(3) present invention is first by Mn-TiO2/g-C3N4It is applied in the preparation of electroluminescent chemiluminescence biosensor, significantly improve the strength and stability of electrogenerated chemiluminescence, substantially increase the detection sensitivity of Electrochemiluminescsensor sensor so that electroluminescent chemiluminescence biosensor achieves the application in real work；The application of this material, is also associated biomolecule sensor, provides Technical Reference such as Optical Electro-Chemistry sensor, electrochemical sensor etc., has the most potential use value.
Detailed description of the invention
Embodiment 1 Mn-TiO2/g-C3N4Preparation
First, take 0.8 mmol manganese salt and join in 5 mL butyl titanates, in whipping process, it is slowly added to 0.5 mL Fluohydric acid., reacts in a kettle. at 160 DEG C 24 hours, after being cooled to room temperature, after ultra-pure water and dehydrated alcohol centrifuge washing three times, it is vacuum dried at 50 DEG C；Secondly, the 150 dried solids of mg and 400 are taken
Mg tripolycyanamide mixes, and grind into powder；Then, putting in Muffle furnace by the powder of grinding, programming rate is 1 DEG C/min, calcines 5 hours at 480 DEG C；Finally, the powder after calcining is cooled to room temperature, i.e. prepares Mn-TiO2/g-C3N4；
Described manganese salt is manganese sulfate.
Embodiment 2 Mn-TiO2/g-C3N4Preparation
First, take 1.0 mmol manganese salt and join in 5 mL butyl titanates, in whipping process, it is slowly added to 0.6 mL Fluohydric acid., reacts in a kettle. at 180 DEG C 21 hours, after being cooled to room temperature, after ultra-pure water and dehydrated alcohol centrifuge washing three times, it is vacuum dried at 50 DEG C；Secondly, the 200 dried solids of mg and 400 are taken
Mg tripolycyanamide mixes, and grind into powder；Then, putting in Muffle furnace by the powder of grinding, programming rate is 2 DEG C/min, calcines 2 hours at 520 DEG C；Finally, the powder after calcining is cooled to room temperature, i.e. prepares Mn-TiO2/g-C3N4；
Described manganese salt is manganese chloride.
Embodiment 3 Mn-TiO2/g-C3N4Preparation
First, take 1.2 mmol manganese salt and join in 5 mL butyl titanates, in whipping process, it is slowly added to 0.8 mL Fluohydric acid., reacts in a kettle. at 200 DEG C 18 hours, after being cooled to room temperature, after ultra-pure water and dehydrated alcohol centrifuge washing three times, it is vacuum dried at 50 DEG C；Secondly, the 250 dried solids of mg and 400 are taken
Mg tripolycyanamide mixes, and grind into powder；Then, putting in Muffle furnace by the powder of grinding, programming rate is 3 DEG C/min, calcines 0.5 hour at 560 DEG C；Finally, the powder after calcining is cooled to room temperature, i.e. prepares Mn-TiO2/g-C3N4；
Described manganese salt is manganese nitrate.
Embodiment 4 The preparation method of electrogenerated chemiluminescence furazolidone biosensor
(1) using a width of 1 cm, a length of 4 cm ITO electro-conductive glass as working electrode, at the Mn-TiO of electrode surface drop coating 8 L2/g-C3N4Colloidal sol, dries under room temperature；
(2) the electrode buffer solution PBS that will obtain in step (1), continues the furazolidone antibody-solutions at electrode surface drop coating 8 L 10 g/mL, preserves and dry in 4 DEG C of refrigerators；
(3) the electrode PBS that will obtain in step (2), continues at the bovine serum albumin solution that electrode surface drop coating 8 L concentration is 100 g/mL, preserves and dry in 4 DEG C of refrigerators；
(4) the electrode PBS that will obtain in step (3), continues the glutaraldehyde solution at electrode surface drop coating 2 L, preserves and dry in 4 DEG C of refrigerators；
(5) the electrode PBS that will obtain in step (4), continues at the horseradish peroxidase solution that electrode surface drop coating 6 L concentration is 20 g/mL, preserves and dry in 4 DEG C of refrigerators；
(6) the electrode PBS that will obtain in step (5), preserves in 4 DEG C of refrigerators after drying, i.e. prepares electrogenerated chemiluminescence furazolidone biosensor；
Described Mn-TiO2/g-C3N4Colloidal sol is by the Mn-TiO of 50 mg2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters, and the hydrosol prepared after ultrasonic 30 min；
Described PBS is the phosphate buffered solution of 10mmol/L, and the pH value of described phosphate buffered solution is 7.4；
Described glutaraldehyde solution be volume ratio be the glutaraldehyde water solution of 2.5%.
Embodiment 5 The preparation method of electrogenerated chemiluminescence furazolidone biosensor
(1) using a width of 1 cm, a length of 4 cm ITO electro-conductive glass as working electrode, at the Mn-TiO of electrode surface drop coating 10 L2/g-C3N4Colloidal sol, dries under room temperature；
(2) the electrode buffer solution PBS that will obtain in step (1), continues the furazolidone antibody-solutions at electrode surface drop coating 10 L 10 g/mL, preserves and dry in 4 DEG C of refrigerators；
(3) the electrode PBS that will obtain in step (2), continues at the bovine serum albumin solution that electrode surface drop coating 10 L concentration is 100 g/mL, preserves and dry in 4 DEG C of refrigerators；
(4) the electrode PBS that will obtain in step (3), continues the glutaraldehyde solution at electrode surface drop coating 3 L, preserves and dry in 4 DEG C of refrigerators；
(5) the electrode PBS that will obtain in step (4), continues at the horseradish peroxidase solution that electrode surface drop coating 8 L concentration is 20 g/mL, preserves and dry in 4 DEG C of refrigerators；
(6) the electrode PBS that will obtain in step (5), preserves in 4 DEG C of refrigerators after drying, i.e. prepares electrogenerated chemiluminescence furazolidone biosensor；
Described Mn-TiO2/g-C3N4Colloidal sol is by the Mn-TiO of 50 mg2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters, and the hydrosol prepared after ultrasonic 30 min；
Described PBS is 10
The phosphate buffered solution of mmol/L, the pH value of described phosphate buffered solution is 7.4；
Described glutaraldehyde solution be volume ratio be the glutaraldehyde water solution of 2.5%.
Embodiment 6 The preparation method of electrogenerated chemiluminescence furazolidone biosensor
(1) using a width of 1 cm, a length of 4 cm ITO electro-conductive glass as working electrode, at the Mn-TiO of electrode surface drop coating 12 L2/g-C3N4Colloidal sol, dries under room temperature；
(2) the electrode buffer solution PBS that will obtain in step (1), continues the furazolidone antibody-solutions at electrode surface drop coating 12 L 10 g/mL, preserves and dry in 4 DEG C of refrigerators；
(3) the electrode PBS that will obtain in step (2), continues at the bovine serum albumin solution that electrode surface drop coating 12 L concentration is 100 g/mL, preserves and dry in 4 DEG C of refrigerators；
(4) the electrode PBS that will obtain in step (3), continues the glutaraldehyde solution at electrode surface drop coating 4 L, preserves and dry in 4 DEG C of refrigerators；
(5) the electrode PBS that will obtain in step (4), continues at the horseradish peroxidase solution that electrode surface drop coating 10 L concentration is 20 g/mL, preserves and dry in 4 DEG C of refrigerators；
(6) the electrode PBS that will obtain in step (5), preserves in 4 DEG C of refrigerators after drying, i.e. prepares electrogenerated chemiluminescence furazolidone biosensor；
Described Mn-TiO2/g-C3N4Colloidal sol is by the Mn-TiO of 50 mg2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters, and the hydrosol prepared after ultrasonic 30 min；
Described PBS is 10
The phosphate buffered solution of mmol/L, the pH value of described phosphate buffered solution is 7.4；
Described glutaraldehyde solution be volume ratio be the glutaraldehyde water solution of 2.5%.
Embodiment 7 The electrogenerated chemiluminescence furazolidone biosensor of embodiment 1 ~ 6 preparation, is applied to the detection of furazolidone, and step is as follows:
(1) standard solution preparation: prepare the furazolidone standard solution of one group of variable concentrations including blank standard specimen；
(2) working electrode is modified: be working electrode by the electrogenerated chemiluminescence furazolidone biosensor prepared by preparation method as claimed in claim 1, by the drop coating respectively of the furazolidone standard solution of the variable concentrations of preparation in step (1) to working electrode surface, 4 DEG C of refrigerators preserve；
(3) working curve is drawn: saturated calomel electrode, be connected on electrochemiluminescdetection detection equipment as reference electrode, platinum electrode as auxiliary electrode, the working electrode composition three-electrode system modified with step (2)；Successively add the K of 15 mL in a cell2S2O8Solution and the H of 100 L2O2Solution；Cyclical voltage is applied with the cyclic voltammetry working electrode to assembling；Relation between light signal strength and the furazolidone antigen concentration of standard solution of the electrogenerated chemiluminescence according to gained, drawing curve；The light signal strength of blank standard specimen is designated asD 0, the light signal strength of the furazolidone standard solution containing variable concentrations is designated asD i, the difference that response light signal intensity reduces is ΔD =D 0-D i, ΔDMass concentration with furazolidone standard solutionCBetween linear, draw ΔD-CWorking curve；The linear detection range of furazolidone is: 0.003 ~ 100 ng/mL, and detection is limited to: 1.1 pg/mL；
(4) detection of furazolidone: replace the furazolidone standard solution in step a with testing sample, detect according to the method in step b and c, the difference DELTA reduced according to response light signal intensityDAnd working curve, obtain the content of furazolidone in testing sample；
Described K2S2O8Solution is by 1 mol K2S2O8Be dissolved in the pH=6.5 buffer solution of 10 L with 1 mol KCl and preparing, described pH=6.5 buffer solution be pH value be the phosphate buffered solution of 6.5；
Described H2O2Solution is aqueous hydrogen peroxide solution, and the concentration of described aqueous hydrogen peroxide solution is 10%.
Claims (2)
1. a preparation method for electrogenerated chemiluminescence furazolidone biosensor, described electrogenerated chemiluminescence furazolidone biosensor is by working electrode, Mn-TiO2/g-C3N4, furazolidone antibody, bovine serum albumin, glutaraldehyde, horseradish peroxidase composition；Described Mn-TiO2/g-C3N4For additive Mn titanium dioxide nanoplate In-situ reaction carbonitride two-dimensional nano composite；
It is characterized in that, described preparation method includes following preparation process:
a. Mn-TiO2/g-C3N4Preparation；
B. the preparation of electrogenerated chemiluminescence furazolidone biosensor；
Wherein, step a prepares Mn-TiO2/g-C3N4Concretely comprise the following steps:
First, take 0.8 ~ 1.2 mmol manganese salt and join in 5 mL butyl titanates, in whipping process, it is slowly added to 0.5 ~ 0.8 mL Fluohydric acid., reacts in a kettle. at 160 ~ 200 DEG C 18 ~ 24 hours, after being cooled to room temperature, after ultra-pure water and dehydrated alcohol centrifuge washing three times, it is vacuum dried at 50 DEG C；Secondly, 150 ~ 250 are taken
The dried solid of mg mixes with 400mg tripolycyanamide, and grind into powder；Then, putting in Muffle furnace by the powder of grinding, programming rate is 1 ~ 3
DEG C/min, calcine 0.5 ~ 5 hour at 480 ~ 560 DEG C；Finally, the powder after calcining is cooled to room temperature, i.e. prepares Mn-TiO2/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 of electrogenerated chemiluminescence furazolidone biosensor:
(1) with ITO electro-conductive glass as working electrode, at the Mn-TiO of electrode surface drop coating 8 ~ 12 L2/g-C3N4Colloidal sol, dries under room temperature；
(2) the electrode buffer solution PBS that will obtain in step (1), continues in electrode surface drop coating 8 ~ 12
The furazolidone antibody-solutions of L 10 g/mL, preserves in 4 DEG C of refrigerators and dries；
(3) the electrode PBS that will obtain in step (2), continues in electrode surface drop coating 8 ~ 12
L concentration is the bovine serum albumin solution of 100 g/mL, preserves and dry in 4 DEG C of refrigerators；
(4) the electrode PBS that will obtain in step (3), continues in electrode surface drop coating 2 ~ 4
The glutaraldehyde solution of L, preserves in 4 DEG C of refrigerators and dries；
(5) the electrode PBS that will obtain in step (4), continues in electrode surface drop coating 6 ~ 10
L concentration is the horseradish peroxidase solution of 20 g/mL, preserves and dry in 4 DEG C of refrigerators；
(6) the electrode PBS that will obtain in step (5), preserves in 4 DEG C of refrigerators after drying, i.e. prepares electrogenerated chemiluminescence furazolidone biosensor；
Described Mn-TiO2/g-C3N4Solution is by the Mn-TiO of 50 mg2/g-C3N4Powder is dissolved in 10 mL ultra-pure waters, and the hydrosol prepared after ultrasonic 30 min；
Described PBS is the phosphate buffered solution of 10mmol/L, and the pH value of described phosphate buffered solution is 7.4；
Described glutaraldehyde solution be volume ratio be the glutaraldehyde water solution of 2.5%.
2. the application of electrogenerated chemiluminescence furazolidone biosensor prepared by preparation method as claimed in claim 1, it is characterised in that include following applying step:
A. standard solution preparation: prepare the furazolidone standard solution of one group of variable concentrations including blank standard specimen；
B. working electrode is modified: be working electrode by the electrogenerated chemiluminescence furazolidone biosensor prepared by preparation method as claimed in claim 1, by the drop coating respectively of the furazolidone standard solution of the variable concentrations of preparation in step a to working electrode surface, 4 DEG C of refrigerators preserve；
C. working curve is drawn:, be connected on electrochemiluminescdetection detection equipment as reference electrode, platinum electrode as to electrode, the working electrode composition three-electrode system modified with step b by Ag/AgCl electrode；Successively add the K of 15 mL in a cell2S2O8Solution and the H of 100 L2O2Solution；Cyclical voltage is applied with the cyclic voltammetry working electrode to assembling；Relation between light signal strength and the furazolidone antigen concentration of standard solution of the electrogenerated chemiluminescence according to gained, drawing curve；The light signal strength of blank standard specimen is designated asD 0, the light signal strength of the furazolidone standard solution containing variable concentrations is designated asD i, the difference that response light signal intensity reduces is ΔD =D 0-D i, ΔDMass concentration with furazolidone standard solutionCBetween linear, draw ΔD-CWorking curve；
D. the detection of furazolidone: replace the furazolidone standard solution in step a with testing sample, detect according to the method in step b and c, the difference DELTA reduced according to response light signal intensityDAnd working curve, obtain the content of furazolidone in testing sample；
Described K2S2O8Solution is by 1 mol K2S2O8Be dissolved in the pH=6.5 buffer solution of 10 L with 1 mol KCl and preparing, described pH=6.5 buffer solution be pH value be the phosphate buffered solution of 6.5；
Described H2O2Solution is aqueous hydrogen peroxide solution, and the concentration of described aqueous hydrogen peroxide solution is 10%.
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|---|---|---|---|---|
| CN107247079A (en) * | 2017-07-19 | 2017-10-13 | 济南大学 | A kind of electrochemiluminescdetection detection method of easy glyphosate pesticide |
| CN110296980A (en) * | 2019-07-16 | 2019-10-01 | 常州大学 | A kind of method of Electrochemiluminescince detection furazolidone |
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| US3980434A (en) * | 1975-07-30 | 1976-09-14 | Morton-Norwich Products, Inc. | Method for determining furazolidone in animal tissue |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107247079A (en) * | 2017-07-19 | 2017-10-13 | 济南大学 | A kind of electrochemiluminescdetection detection method of easy glyphosate pesticide |
| CN110296980A (en) * | 2019-07-16 | 2019-10-01 | 常州大学 | A kind of method of Electrochemiluminescince detection furazolidone |
| CN110296980B (en) * | 2019-07-16 | 2022-02-11 | 常州大学 | Method for detecting furazolidone by electrochemical luminescence method |
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|---|---|
| CN105717099B (en) | 2018-03-30 |
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