CN105548302B - A kind of detection method to radical reaction - Google Patents
A kind of detection method to radical reaction Download PDFInfo
- Publication number
- CN105548302B CN105548302B CN201610069328.8A CN201610069328A CN105548302B CN 105548302 B CN105548302 B CN 105548302B CN 201610069328 A CN201610069328 A CN 201610069328A CN 105548302 B CN105548302 B CN 105548302B
- Authority
- CN
- China
- Prior art keywords
- radical reaction
- electrode
- detection method
- detection
- sensitivity
- 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.)
- Active
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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to analysis and testing technology fields, and in particular to a kind of detection method to radical reaction.Using by radical reaction polymerizing cationically or anionic monomer, using the polymer membrane electrode to polycation or polyanion sensitivity as indicator electrode, and then to the detection of radical reaction and the substance that radical reaction can be triggered.The method of the present invention high sensitivity, the detection available for a variety of chemistry and biological targets object.
Description
Technical field
The invention belongs to analysis and testing technology fields, and in particular to a kind of detection method to radical reaction.
Background technology
Radical reaction is widely present in a variety of chemistry and biological process.Because it is generated through Raolical polymerizable
Polymer lacks signal reports function, so traditional detection method to radical reaction needs that reaction product is marked
Or it modifies to complete.This mark or modification generally require longer reaction time and cumbersome operating process, and
This kind of method easily disturbed in by sample by turbidity and colourity, is unfavorable for promoting and applying.
Polymer film electric potential type sensing technology is of simple structure and low cost, is disturbed from turbidity and colourity, in many fields
With irreplaceable advantage.Although a variety of biological targets are widely used in the polymer membrane electrode of polyion sensitivity
The detection of object, but the report for using it for artificial synthesized polyion detection is relatively fewer.
The content of the invention
Detection method and its application it is an object of the invention to provide a kind of radical reaction.
To achieve the above object, the technical solution adopted by the present invention is：
A kind of detection method to radical reaction, using by radical reaction polymerizing cationically or anionic monomer,
Using the polymer membrane electrode to polycation or polyanion sensitivity as indicator electrode, and then to radical reaction and can trigger certainly
By the detection of the substance of base reaction.
The cationic monomer be may occur from it is by base polymerisation and positively charged in the case where acidity is neutrallty condition
Substance；
The anionic monomer be may occur from it is by base polymerisation and negatively charged in the case where acidity is neutrallty condition
Substance.
The cationic monomer is N- propyl methacrylamides, N- aminoethylmethacrylamides, N- ammonia
Base butyl methyl acrylamide, N- aminoethyl acrylamides, N- aminopropyl acrylamides, N- aminobutyls acrylamide or
Acrylamide；
The anionic monomer is methacrylic acid, acrylic acid, methylcrotonic acid, butenoic acid or ethylacrylic acid.
The polymer membrane electrode to polycation sensitivity is the electrode doped with cation-exchanger；Cation is handed over
Agent is changed as four (4- fluorophenyls) Boratexes, four (4- chlorphenyls) Boratexes, four (4- aminomethyl phenyls) Boratexes, four (4- fluoroforms
Base phenyl) Boratex, four (3,5- bis- (trifluoromethyl) phenyl) Boratexes or sodium tetraphenylborate.
The polymer membrane electrode to polyanion sensitivity is the electrode doped with anionite；Anion is handed over
Agent is changed as dinonylnaphthalene sulfonic acid, dinonylnaphthalene sulfonic acid sodium, two naphthalene sulfonic acids of dinonyl, dinonyl sodium dinaphthalenesulfonate, dotriacontyl
Monomethyl ammonium chloride, dotetracontane ammonium chloride, dotriacontyl monomethyl ammonium bromide or dotetracontane base ammonium bromide.
Specific detection method：
(1) it is as working electrode, saturated calomel electrode using the polymer membrane electrode to polycation or polyanion sensitivity
Reference electrode carries out potentiometric detection；
(2) addition and cation or anionic monomer in 50mM HEPES (pH=7.4) solution, add in various concentration
Determinand triggers free responding；
(3) working electrode is put into record initial potential value in 50mM HEPES (pH=7.4) solution, then by step
(2) determinand for entering various concentration triggers the product of free responding to add in record difference in 50mM HEPES (pH=7.4) solution
The changing value for the polyion current potential that determinand generates；
(4) for the electric potential signal of the polyion generated using different determinands as ordinate, concentration draws standard work for abscissa
Make curve；
(5) actual sample is tested according to the step of (1)-(4), is calculated according to standard working curve to be measured in sample
The concentration value of object.
The method of the present invention can be used for a variety of target analyte detections for triggering radical reaction, such as horseradish peroxidase, sweet smell
Pause reagent, G- tetrads nuclease, catalase, laccase.
Advantage for present invention：The present invention is using radical reaction polymerizing cationically or anionic monomer, with to poly-
Cation or the polymer membrane electrode of polyanion sensitivity are indicator electrode, it can be achieved that radical reaction with that can trigger free radical
The detection of the substance of reaction.This method is easy to operate, high sensitivity, is not required to cumbersome mark and modification step；This method can use
In a variety of object such as horseradish peroxidase, Fenton reagent, G- tetrads nuclease, peroxidating for triggering radical reaction
The detection of hydrogen enzyme, laccase etc..
Description of the drawings
Fig. 1 is that the potential response figure (a) of horseradish peroxidase and its calibration curve (b) are detected in the embodiment of the present invention 1.
Fig. 2 is to detect Fe in the embodiment of the present invention 22+And Cu2+Calibration curve.
Fig. 3 is that the potential response figure (a) of catalase and its calibration curve (b) are detected in the embodiment of the present invention 3.
Fig. 4 is the potential response figure (a) and its calibration curve (b) that G- tetrad nucleases are detected in the embodiment of the present invention 4.
Specific embodiment
Specific examples below is used for further illustrating the present invention, but the present invention is only limitted to absolutely not these examples.
The present invention is using radical reaction polymerizing cationically or anionic monomer, with sensitive to polycation or polyanion
Polymer membrane electrode be indicator electrode, it can be achieved that detection to radical reaction and the substance that radical reaction can be triggered.
Embodiment 1
Using N- propyl methacrylamides as cationic monomer, with the polymer membrane electrode of dinonylnaphthalene sulfonic acid doping
For indicator electrode, the detection to horseradish peroxidase is realized.Its operating procedure is as follows：
(1) preparation of polymer membrane electrode
It weighs 196mg polyvinyl chloride, 196mg ortho-nitrophenyls octyl ether, 4.0mg dinonylnaphthalene sulfonic acids and is dissolved in 3.0mL tetrahydrochysene furans
In muttering, poured into after stirring evenly in the glass ring that fixed internal diameter on a glass is 3.5cm.Treat that tetrahydrofuran volatilization finishes
Afterwards, film is cut into sequin that internal diameter is 3mm using card punch and pastes bottom and be cased in the liquid-transfering gun pipette tips of pvc pipe.
The HEPES solution used is as internal-filling liquid and activating solution activated electrode, for use.
(2) initiation of radical reaction
Use addition glucose oxidase (200nM), glucose in 50mM HEPES buffer solutions (pH=7.4)
(10mM), N- propyl methacrylamides (0.5M), acetylacetone,2,4-pentanedione (0.5mM) and various concentration (5 × 10-7、1×10-6、
3×10-6、5×10-6、1×10-5U/mL horseradish peroxidase), reacts half an hour at room temperature.
(3) acquisition of electric potential signal
Working electrode is put into record initial potential value in 50mM HEPES (pH=7.4) solution, is then added in above-mentioned anti-
The product answered makes 3300 times of the product dilution of above-mentioned reaction, records the changing value of current potential.
(4) drafting of standard working curve
For the electric potential signal of the polyion generated using different determinands as ordinate, concentration draws standard work song for abscissa
Line, experimental result are shown in Fig. 1.
(5) actual sample is tested according to the step of (1)-(4), is calculated according to standard working curve to be measured in sample
The concentration value of object.
Can realize highly sensitive detection to horseradish peroxidase using the sensor as seen from Figure 1, the range of linearity for 1 ×
10-5~5 × 10-7U/mL, detection are limited to 2 × 10-7U/mL。
Embodiment 2
The present invention is using N- propyl methacrylamides as cationic monomer, with the polymer of dinonylnaphthalene sulfonic acid doping
Membrane electrode is indicator electrode, realizes Fenton reagent Fe2+And Cu2+Detection.Its operating procedure is as follows：
(1) preparation of polymer membrane electrode
It weighs 196mg polyvinyl chloride, 196mg ortho-nitrophenyls octyl ether, 4.0mg dinonylnaphthalene sulfonic acids and is dissolved in 3.0mL tetrahydrochysene furans
In muttering, poured into after stirring evenly in the glass ring that fixed internal diameter on a glass is 3.5cm.Treat that tetrahydrofuran volatilization finishes
Afterwards, film is cut into sequin that internal diameter is 3mm using card punch and pastes bottom and be cased in the liquid-transfering gun pipette tips of pvc pipe.
The HEPES solution used is as internal-filling liquid and activating solution activated electrode, for use.
(2) initiation of radical reaction
Use addition glucose oxidase (200nM), glucose in 50mM HEPES buffer solutions (pH=7.4)
(10mM), N- propyl methacrylamides (0.5M), acetylacetone,2,4-pentanedione (0.5mM) and various concentration (0.5,1,2,5,
10ppb)Fe2+(0.1,0.5,1,2,5ppb) Cu2+, half an hour is reacted at room temperature.
(3) acquisition of electric potential signal
Working electrode is put into record initial potential value in 50mM HEPES (pH=7.4) solution, is then added in above-mentioned anti-
The product answered makes 3300 times of the product dilution of above-mentioned reaction, records the changing value of current potential.
(4) drafting of standard working curve
For the electric potential signal of the polyion generated using different determinands as ordinate, concentration draws standard work song for abscissa
Line, experimental result are shown in Fig. 2.
(5) actual sample is tested according to the step of (1)-(4), is calculated according to standard working curve to be measured in sample
The concentration value of object.
As seen from Figure 2 Fe can be realized using the sensor2+And Cu2+Highly sensitive detection.
Embodiment 3
The present invention is using N- propyl methacrylamides as cationic monomer, with the polymer of dinonylnaphthalene sulfonic acid doping
Membrane electrode is indicator electrode, realizes the detection to catalase.Its operating procedure is as follows：
(1) preparation of polymer membrane electrode
It weighs 196mg polyvinyl chloride, 196mg ortho-nitrophenyls octyl ether, 4.0mg dinonylnaphthalene sulfonic acids and is dissolved in 3.0mL tetrahydrochysene furans
In muttering, poured into after stirring evenly in the glass ring that fixed internal diameter on a glass is 3.5cm.Treat that tetrahydrofuran volatilization finishes
Afterwards, film is cut into sequin that internal diameter is 3mm using card punch and pastes bottom and be cased in the liquid-transfering gun pipette tips of pvc pipe.
The HEPES solution used is as internal-filling liquid and activating solution activated electrode, for use.
(2) initiation of radical reaction
Use addition glucose oxidase (200nM), glucose in 50mM HEPES buffer solutions (pH=7.4)
(the 2 × 10 of (10mM), N- propyl methacrylamides (0.5M), acetylacetone,2,4-pentanedione (2.0mM) and various concentration-7、5×10-7、1×10-6、2×10-6、5×10-6U/mL) catalase reacts half an hour at room temperature.
(3) acquisition of electric potential signal
Working electrode is put into record initial potential value in 50mM HEPES (pH=7.4) solution, is then added in above-mentioned anti-
The product answered makes 3300 times of the product dilution of above-mentioned reaction, records the changing value of current potential.
(4) drafting of standard working curve
For the electric potential signal of the polyion generated using different determinands as ordinate, concentration draws standard work song for abscissa
Line, experimental result are shown in Fig. 3.
(5) actual sample is tested according to the step of (1)-(4), is calculated according to standard working curve to be measured in sample
The concentration value of object.
From the figure 3, it may be seen that can realize the highly sensitive detection to catalase using the sensor, the range of linearity is 5 × 10-6
~2 × 10-7U/mL, detection are limited to 1 × 10-7U/mL。
Embodiment 4
The present invention is using methacrylic acid as anionic monomer, with the polymer film of dotriacontyl monomethyl ammonium chloride doping
Electrode is indicator electrode, realizes the detection to G- tetrad nucleases.Its operating procedure is as follows：
(1) preparation of polymer membrane electrode
196mg polyvinyl chloride, 196mg ortho-nitrophenyls octyl ether, 4.0mg dotriacontyl monomethyl ammonium chlorides is weighed to be dissolved in
In 3.0mL tetrahydrofurans, poured into after stirring evenly in the glass ring that fixed internal diameter on a glass is 3.5cm.Treat tetrahydrochysene furan
It mutters after volatilization, film is cut into sequin that internal diameter is 3mm using card punch and pastes the liquid relief that bottom is cased with pvc pipe
In rifle pipette tips.The HEPES solution used is as internal-filling liquid and activating solution activated electrode, for use.
(2) initiation of radical reaction
Use addition glucose oxidase (200nM), glucose in 50mM HEPES buffer solutions (pH=7.4)
(the 5 × 10 of (10mM), methacrylic acid (0.5M), acetylacetone,2,4-pentanedione (0.5mM) and various concentration-9、1×10-9、5×10-10、1
×10-10、5×10-11M) G- tetrads/ferroheme, reacts half an hour at room temperature.
(3) acquisition of electric potential signal
Working electrode is put into record initial potential value in 50mM HEPES (pH=7.4) solution, is then added in above-mentioned anti-
The product answered makes 4300 times of the product dilution of above-mentioned reaction, records the changing value of current potential.
(4) drafting of standard working curve
For the electric potential signal of the polyion generated using different determinands as ordinate, concentration draws standard work song for abscissa
Line, experimental result are shown in Fig. 4.
(5) actual sample is tested according to the step of (1)-(4), is calculated according to standard working curve to be measured in sample
The concentration value of object.
As shown in Figure 4, highly sensitive detection to G- tetrad nucleases, the range of linearity 5 can be realized using the sensor
×10-9~5 × 10-11M, detection are limited to 2 × 10-11M。
Claims (5)
1. a kind of detection method to radical reaction, it is characterised in that：Using passing through radical reaction polymerizing cationically or the moon
Ion monomer, using the polymer membrane electrode to polycation or polyanion sensitivity as indicator electrode, and then to radical reaction
Detection with the substance that can trigger radical reaction；
Specially：（1）Using the polymer membrane electrode to polycation or polyanion sensitivity as working electrode, saturated calomel electrode
Potentiometric detection is carried out for reference electrode；
（2）Cation or anionic monomer are added in 50 mM HEPES solution, the determinand for adding in various concentration triggers freely
Reaction；
（3）Working electrode is put into record initial potential value in 50 mM HEPES solution, then by step（2）Enter various concentration
Determinand the product of free responding is triggered to add in 50 mM HEPES solution record the polyion current potential that different determinands generate
Changing value；
（4）For the electric potential signal of the polyion generated using different determinands as ordinate, concentration draws standard work song for abscissa
Line；
（5）According to（1）-（4）The step of actual sample is tested, determinand in sample is calculated according to standard working curve
Concentration value.
2. by the detection method of radical reaction described in claim 1, it is characterised in that：
The cationic monomer is that may occur from by base polymerisation and the substance positively charged in the case where acidity is neutrallty condition；
The anionic monomer is that may occur from by base polymerisation and the substance negatively charged in the case where acidity is neutrallty condition.
3. the detection method of the radical reaction as described in claim 2, it is characterised in that：
The cationic monomer is N- propyl methacrylamides, N- aminoethylmethacrylamides, N- amino fourths
Butylmethacrylamide, N- aminoethyl acrylamides, N- aminopropyl acrylamides, N- aminobutyls acrylamide or propylene
Amide；
The anionic monomer is methacrylic acid, acrylic acid, methylcrotonic acid, butenoic acid or ethylacrylic acid.
4. by the detection method of radical reaction described in claim 1, it is characterised in that：
The polymer membrane electrode to polycation sensitivity is the electrode doped with cation-exchanger；Cation-exchanger
For four (4- fluorophenyls) Boratexes, four (4- chlorphenyls) Boratexes, four (4- aminomethyl phenyls) Boratexes, four (4- trifluoromethylbenzenes
Base) Boratex, four (3,5- bis- (trifluoromethyl) phenyl) Boratexes or sodium tetraphenylborate.
5. by the detection method of radical reaction described in claim 1, it is characterised in that：
The polymer membrane electrode to polyanion sensitivity is the electrode doped with anionite；Anionite
For dinonylnaphthalene sulfonic acid, dinonylnaphthalene sulfonic acid sodium, two naphthalene sulfonic acids of dinonyl, dinonyl sodium dinaphthalenesulfonate, one first of dotriacontyl
Ammonium chloride, dotetracontane ammonium chloride, dotriacontyl monomethyl ammonium bromide or dotetracontane base ammonium bromide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610069328.8A CN105548302B (en) | 2016-02-01 | 2016-02-01 | A kind of detection method to radical reaction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610069328.8A CN105548302B (en) | 2016-02-01 | 2016-02-01 | A kind of detection method to radical reaction |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105548302A CN105548302A (en) | 2016-05-04 |
| CN105548302B true CN105548302B (en) | 2018-05-25 |
Family
ID=55827638
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610069328.8A Active CN105548302B (en) | 2016-02-01 | 2016-02-01 | A kind of detection method to radical reaction |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105548302B (en) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100595577C (en) * | 2007-07-23 | 2010-03-24 | 南京工业大学 | Preparation method of enzyme electrode for detecting glucose |
| CN104931563B (en) * | 2015-05-29 | 2017-09-29 | 中国科学院烟台海岸带研究所 | A kind of electric potential type glucose sensor of high selectivity |
-
2016
- 2016-02-01 CN CN201610069328.8A patent/CN105548302B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN105548302A (en) | 2016-05-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lahcen et al. | Label-free electrochemical sensor based on spore-imprinted polymer for Bacillus cereus spore detection | |
| Rkik et al. | Electrochemical determination of levofloxacin antibiotic in biological samples using boron doped diamond electrode | |
| Alizadeh et al. | Selective determination of chloramphenicol at trace level in milk samples by the electrode modified with molecularly imprinted polymer | |
| Liu et al. | Electrochemical microfluidic chip based on molecular imprinting technique applied for therapeutic drug monitoring | |
| Alizadeh et al. | A new molecularly imprinted polymer (MIP)-based electrochemical sensor for monitoring 2, 4, 6-trinitrotoluene (TNT) in natural waters and soil samples | |
| Pushpanjali et al. | The electrochemical resolution of ciprofloxacin, riboflavin and estriol using anionic surfactant and polymer‐modified carbon paste electrode | |
| Giribabu et al. | Nanomolar determination of 4-nitrophenol based on a poly (methylene blue)-modified glassy carbon electrode | |
| Prathish et al. | Molecularly imprinted polymer-based potentiometric sensor for degradation product of chemical warfare agents: Part I. Methylphosphonic acid | |
| Tadi et al. | Electrochemical detection of sulfanilamide using pencil graphite electrode based on molecular imprinting technology | |
| Rius-Ruiz et al. | Solid-state reference electrodes based on carbon nanotubes and polyacrylate membranes | |
| Prasad et al. | Ion imprinted polymer based ion-selective electrode for the trace determination of dysprosium (III) ions | |
| Ağın | Electrochemical determination of amoxicillin on a poly (acridine orange) modified glassy carbon electrode | |
| Guerreiro et al. | New sensing materials of molecularly-imprinted polymers for the selective recognition of chlortetracycline | |
| Li et al. | A ratiometry-induced successive reusable electrochemical aptasensing platform: Efficient monitoring of aflatoxin B1 in peanut | |
| Katsu et al. | Thallium (I)-selective membrane electrodes based on calix [6] arene or calix [5] arene derivatives | |
| Yasri et al. | Chronoamperometric determination of lead ions using PEDOT: PSS modified carbon electrodes | |
| Arvand et al. | Sulfamethoxazole‐imprinted polymeric receptor as ionophore for potentiometric transduction | |
| Brycht et al. | Ultra trace level determination of fenoxanil by highly sensitive square wave adsorptive stripping voltammetry in real samples with a renewable silver amalgam film electrode | |
| Libansky et al. | Construction of an electrochemical cell system based on carbon composite film electrodes and its application for voltammetric determination of triclosan | |
| Lee et al. | Urea-functionalized calix [4] arenes as carriers for carbonate-selective electrodes | |
| Ali et al. | Development of novel potentiometric sensors for determination of lidocaine hydrochloride in pharmaceutical preparations, serum and urine samples | |
| CN102565174A (en) | Ionic liquid polyaniline modified electrode-electro chemical analysis system | |
| Martin et al. | Solid‐contact Acetate‐selective Electrode Based on a 1, 3‐bis (carbazolyl) urea‐ionophore | |
| Lou et al. | A novel electrochemical sensing platform for anions based on conducting polymer film modified electrodes integrated on paper-based chips | |
| Zaabal et al. | A facile strategy for construction of sensor for detection of ondansetron and investigation of its redox behavior and thermodynamic parameters |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |