CN108132290A - A kind of electrochemica biological sensor and its preparation and application based on gold nano triangular plate and horseradish peroxidase modified electrode - Google Patents
A kind of electrochemica biological sensor and its preparation and application based on gold nano triangular plate and horseradish peroxidase modified electrode Download PDFInfo
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- CN108132290A CN108132290A CN201810100109.0A CN201810100109A CN108132290A CN 108132290 A CN108132290 A CN 108132290A CN 201810100109 A CN201810100109 A CN 201810100109A CN 108132290 A CN108132290 A CN 108132290A
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- 108010001336 Horseradish Peroxidase Proteins 0.000 title claims abstract description 85
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 title claims abstract description 24
- 239000010931 gold Substances 0.000 title claims abstract description 24
- 229910052737 gold Inorganic materials 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 229920000557 Nafion® Polymers 0.000 claims abstract description 58
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 29
- 238000001548 drop coating Methods 0.000 claims abstract description 15
- 238000001035 drying Methods 0.000 claims abstract description 3
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 claims description 24
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 22
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 15
- 238000000576 coating method Methods 0.000 claims description 12
- 230000009467 reduction Effects 0.000 claims description 12
- 238000006555 catalytic reaction Methods 0.000 claims description 10
- 229910052799 carbon Inorganic materials 0.000 claims description 9
- 230000005611 electricity Effects 0.000 claims description 7
- 239000002608 ionic liquid Substances 0.000 claims description 7
- 239000011248 coating agent Substances 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 235000010288 sodium nitrite Nutrition 0.000 claims description 6
- 230000003287 optical effect Effects 0.000 claims description 5
- 239000000758 substrate Substances 0.000 claims description 5
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 claims description 4
- 150000001721 carbon Chemical class 0.000 claims description 4
- 239000004570 mortar (masonry) Substances 0.000 claims description 4
- 238000001514 detection method Methods 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- -1 N- hexyl pyridine hexafluorophosphates Chemical class 0.000 claims description 2
- 238000003801 milling Methods 0.000 claims description 2
- 239000000243 solution Substances 0.000 description 25
- 235000019441 ethanol Nutrition 0.000 description 8
- 238000002484 cyclic voltammetry Methods 0.000 description 6
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 208000005374 Poisoning Diseases 0.000 description 4
- 231100000572 poisoning Toxicity 0.000 description 4
- 230000000607 poisoning effect Effects 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 235000011330 Armoracia rusticana Nutrition 0.000 description 3
- 240000003291 Armoracia rusticana Species 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 3
- 230000003197 catalytic effect Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000005518 electrochemistry Effects 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 108010061951 Methemoglobin Proteins 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 238000004847 absorption spectroscopy Methods 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000010439 graphite Substances 0.000 description 2
- 229910002804 graphite Inorganic materials 0.000 description 2
- 238000001453 impedance spectrum Methods 0.000 description 2
- 238000012417 linear regression Methods 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 235000013622 meat product Nutrition 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000008055 phosphate buffer solution Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000027756 respiratory electron transport chain Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000003796 beauty Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 239000000645 desinfectant Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000011263 electroactive material Substances 0.000 description 1
- 238000003487 electrochemical reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 150000003278 haem Chemical group 0.000 description 1
- 230000026030 halogenation Effects 0.000 description 1
- 238000005658 halogenation reaction Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 231100000567 intoxicating Toxicity 0.000 description 1
- 230000002673 intoxicating effect Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000002082 metal nanoparticle Substances 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- ZAOCWQZQPKGTRN-UHFFFAOYSA-N nitrous acid;sodium Chemical compound [Na].ON=O ZAOCWQZQPKGTRN-UHFFFAOYSA-N 0.000 description 1
- 229910000510 noble metal Inorganic materials 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 150000002896 organic halogen compounds Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 238000010422 painting Methods 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000002468 redox effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 235000020681 well water Nutrition 0.000 description 1
- 239000002349 well water Substances 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
Classifications
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- 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
-
- 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
- G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
-
- 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
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Electrochemistry (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to a kind of electrochemica biological sensor based on gold nano triangular plate and horseradish peroxidase modified electrode and its preparation and application；The preparation method based on gold nano triangular plate and horseradish peroxidase modified electrode, includes the following steps：(1) appropriate AuNTs solution drop coating is taken to be protected from light naturally dry under room temperature, obtain AuNTs/CILE electrodes on CILE basal electrodes surface；(2) appropriate HRP solution drop coating is taken to be protected from light naturally dry at room temperature, obtain HRP/AuNTs/CILE electrodes in AuNTs/CILE electrode surfaces；(3) appropriate Nafion ethanol solutions drop coating is taken in HRP/AuNTs/CILE electrode surfaces, is protected from light at room temperature after drying to get to Nafion/HRP/AuNTs/CILE electrodes.
Description
Technical field
The invention belongs to electrochemica biological sensor fields, and in particular to one kind is based on gold nano triangular plate and horseradish peroxide
The electrochemica biological sensor of compound enzyme modified electrode and its preparation and application.
Background technology
In recent years, aspherical noble metal nano particles are because of the uniqueness of its structure and properties, more by researcher
Concern wherein.The unique physicochemical property of gold nano-material makes it with more superior sensitivity, stability and selectivity it is new
It has a wide range of applications in terms of type sensing strategy.Nanogold particle as a sensing element can be used for detect metal from
The various targets such as son, small molecule, protein, nucleotide, cell become a kind of novel, rapid, efficient means.Gold nano three
Cornual plate structure have anisotropic surface can with surface plasma body resonant vibration characteristic, in sensing, optical material, catalysis and biology
There is important application in field.In a variety of anisotropy nano particles, gold and Nano silver piece (such as triangular plate and hexagon piece)
Lateral dimension is much larger than thickness, therefore with more special property.Compared to Nano silver piece, gold-nano-piece has higher chemistry
Stability.
Horseradish peroxidase (HRP) is combined by zymoprotein and activated centre ferriporphyrin, is a kind of water soluble oxidized
Reduced form glycoprotein, is widely present in plant, the content highest in horseradish.HRP relative molecular masses are about 40000, peptide
Chain is coiled into a three-dimensional macro structure for being similar to elliposoidal.
Trichloroacetic acid (TCA) is used as a kind of organohalogen compounds, is the by-product of the metabolism of carbon tetrachloride, has and stablize
The characteristics of property is strong, it is difficult to halogenation be gone by direct-reduction on general electrode.Being widely used for it is general, and agricultural aspect can be used as and remove
Careless agent, medical and beauty treatment aspect can be used as wrinkle removing agent and agent of tatooing, used also as the disinfectant of drinking water, TCA in daily life
The lung neoplasm in organism is can result in, cancer can also be caused and influences human reproduction's function.The World Health Organization clearly advises
The security presence amount for determining trichloroacetic acid in water environment is 100 μ g L-1。
Nitrite is white or yellowish rhomboidal crystal, soluble easily in water to be slightly soluble in methanol, ethyl alcohol, ether in liquefied ammonia,
Hygroscopicity is strong, is usually used in common a kind of food in the mordant of textile dyeing and meat products and other food processing process
Additive.It, which has, inhibits meat poisoning fusiform bacillus, makes meat color development and enhances the effect of flavor.But people eats 0.3g
Poisoning symptom may occur to 0.5g, if disposably eating more than 3g by mistake, it is possible to cause death.Nitrite poisoning is
If it is more than national standard to refer to its content in food, due to eat nitrate or the higher marinated meat products of content of nitrite,
Pickles and rotten vegetables cause poisoning or accidentally industrial sodium nitrite are eaten as salt and is caused, also seen in drink
With the intoxicating phenomenon after the bitter well water containing nitrate or nitrite, steamer water.Since nitrite can make in blood normally
The low Ferri-hemoglobin of oxygen carrying is oxidized to ferrihemoglobin, and then loses oxygen carrying capacity and cause histanoxia, leads to body
It changes.
Invention content
The present invention provides a kind of based on gold nano triangular plate and horseradish peroxidase modified electrode, it is characterised in that the electricity
Pole includes basal electrode, gold nano triangular plate (AuNTs) coating, horseradish peroxidase (HRP) coating, Nafion coatings, institute
Basal electrode is stated as Ionic Liquid Modified carbon paste electrode (CILE).
The present invention provides one kind and is followed successively by CILE basal electrodes, AuNTs coatings, HRP coatings, Nafion paintings from inside to outside
The electrode of layer, i.e. Nafion/HRP/AuNTs/CILE electrodes.
The present invention provides a kind of electrochemica biological sensor part, it is characterised in that is based on gold nano triangle so that the present invention is above-mentioned
Piece and horseradish peroxidase modified electrode are as working electrode, preferably using Nafion/HRP/AuNTs/CILE electrodes as work
Make electrode.
Another embodiment of the present invention provides above-mentioned based on gold nano triangular plate and horseradish peroxidase modified electrode
The application of (preferably Nafion/HRP/AuNTs/CILE electrodes) in electrochemica biological sensor part is prepared.
Another embodiment of the present invention provides above-mentioned based on gold nano triangular plate and horseradish peroxidase modified electrode
The preparation method of (preferably Nafion/HRP/AuNTs/CILE electrodes), it is characterised in that include the following steps：
(1) appropriate AuNTs solution drop coating is taken to be protected from light naturally dry under room temperature, obtain on CILE basal electrodes surface
AuNTs/CILE electrodes；
(2) appropriate HRP solution drop coating is taken to be protected from light naturally dry at room temperature, obtain in AuNTs/CILE electrode surfaces
HRP/AuNTs/CILE electrodes；
(3) appropriate Nafion ethanol solutions drop coating is taken to be protected from light at room temperature after drying in HRP/AuNTs/CILE electrode surfaces,
Obtain Nafion/HRP/AuNTs/CILE electrodes；
The preparation method of step (1) the CILE basal electrodes includes the following steps：By suitable graphite powder, N- hexyl pyrroles
Pyridine hexafluorophosphate HPPF6, atoleine be placed in mortar and grind uniform, obtain carbon paste, carbon paste then inserted into glass electrode
It is compacted in pipe, interpolation copper wire obtains Ionic Liquid Modified carbon paste electrode, i.e. CILE basal electrodes as conducting wire；Wherein, graphite powder
With HPPF6Mass ratio be 1.5~2.5:1, preferably 2:1；Every gram of HPPF6Use 500~700 μ L atoleines, preferably 500 μ L；
Milling time is 3.0-5.0h；Glass electrode bore is 4mm；
The optical density (OD) of AuNTs solution described in step (1) be 1, dosage using can even spread CILE electrode surfaces as
Preferably；
A concentration of 10~20mgmL of step (2) the HRP solution-1, preferably 15mgmL-1, dosage with can uniformly
Coating AuNTs/CILE electrode surfaces are advisable；
The volumetric concentration of step (3) the Nafion ethanol solutions is 0.3~0.7%, preferably 0.5%, and dosage is with energy
Even spread HRP/AuNTs/CILE electrode surfaces are advisable.
Another embodiment of the present invention provides above-mentioned based on gold nano triangular plate and horseradish peroxidase modified electrode
The application of (preferably Nafion/HRP/AuNTs/CILE electrodes) in electrochemica biological sensor part is prepared.
Another embodiment of the present invention provides above-mentioned based on gold nano triangular plate and horseradish peroxidase modified electrode
The application of (preferably Nafion/HRP/AuNTs/CILE electrodes) in catalysis restores trichloroacetic acid.
Another embodiment of the present invention provides above-mentioned based on gold nano triangular plate and horseradish peroxidase modified electrode
The application of (preferably Nafion/HRP/AuNTs/CILE electrodes) in trichloroacetic acid is detected.
Another embodiment of the present invention provides above-mentioned based on gold nano triangular plate and horseradish peroxidase modified electrode
The application of (preferably Nafion/HRP/AuNTs/CILE electrodes) in catalysis restores sodium nitrite.
Another embodiment of the present invention provides above-mentioned based on gold nano triangular plate and horseradish peroxidase modified electrode
The application of (preferably Nafion/HRP/AuNTs/CILE electrodes) in sodium nitrite is detected.
The use of AuNTs solution, HRP solution, Nafion ethanol solutions in above-mentioned preparation method step (1)-(3) of the present invention
Amount, with can even spread electrode surface be advisable, those skilled in the art can according to electrode and be coated with solution property, rationally
Select the dosage of AuNTs solution, HRP solution, Nafion ethanol solutions；Nafion ethanol solutions can be matched with Du Pont D520 type solution
System.
In the present invention, CILE is Ionic Liquid Modified carbon paste electrode, and AuNTs is gold nano triangular plate, and HRP is horseradish mistake
Oxide enzyme, Nafion are perfluorinated sulfonic acid-teflon-copolymers, and TCA is trichloroacetic acid, NaNO2For sodium nitrite, PBS
Buffer solution is phosphate buffer solution；Be related to solution is aqueous solution (except having indicated).
The electrocatalytic reaction of gold nano triangular plate and horseradish peroxidase modified electrode of the present invention is in pH 3.0
Phosphate buffer solution in carry out.
Compared with prior art, the advantage of the invention is that：(1) a kind of novel third generation electrochemistry is prepared in the present invention
Enzyme sensor part Nafion/HRP/AuNTs/CILE electrodes, the electrode not only maintain original conformation of HRP, show a pair
Symmetrical redox peaks, and since the addition of AuNTs so that impedance value is smaller, in terms of redox property and electric conductivity
Better than Nafion/CILE, AuNTs/CILE, Nafion/AuNTs/CILE, Nafion/HRP/CILE electrode；(2) it is of the invention
Nafion/HRP/AuNTs/CILE electrodes overcome the problems of preceding two generations enzyme sensor, to trichloroacetic acid and nitrous acid
Sodium has Well-recovered, can effectively amplify current signal, the detection range of linearity is wide, and detection limits low, high sensitivity.
Description of the drawings
Fig. 1：The transmission electron microscope picture of AuNTs
Fig. 2：(a) ultraviolet-visible absorption spectroscopy figure of HRP and (b) HRP-AuNTs mixed solutions.
Fig. 3：Different modifying electrode is in the PBS buffer solutions of pH 3.0, sweep speed 100mV s-1Cyclic voltammogram,
In：(a) Nafion/CILE, (b) Nafion/AuNTs/CILE, (c) Nafion/HRP/CILE and (d) Nafion/HRP/
AuNTs/CILE。
Fig. 4：The electrochemical AC impedance collection of illustrative plates of different modifying electrode, (a) AuNTs/CILE, (b) Nafion/HRP/
AuNTs/CILE, (c) Nafion/CILE and (d) Nafion/HRP/CILE.
Fig. 5：Modified electrode in the presence of various concentration TCA cyclic voltammogram (a to k be 0.0,30.0,60.0,90.0,
120.0,150.0,180.0,210.0,240.0,260.0,300.0mmol L-1) (illustration：Reduction peak current and TCA concentration it
Between relation curve).
Fig. 6：Modified electrode is in various concentration NaNO2In the presence of cyclic voltammogram (a to l be 0.0,1.0,3.0,5.0,
10.0,20.0,30.0,42.0,50.0,62.0,70.0,80.0mmol L-1) (illustration：Reduction peak current and NaNO2Concentration it
Between relation curve).
Specific embodiment
For the ease of a further understanding of the present invention, examples provided below has done more detailed description to it.But
It is that these embodiments only are not used for limiting the scope of the present invention or implementation principle, reality of the invention for being better understood from inventing
The mode of applying is not limited to the following contents.
1 transmission electron microscope picture of embodiment
Used AuNTs is characterized with transmission electron microscope, the results are shown in Figure 1, shows as the gold-nano-piece of equilateral triangle,
Close to equilateral, three angles are fillet on three sides, and particle size is the nanometer of a standard with center to three vertex about 60nm
Triangle.
2 ultraviolet-visible absorption spectroscopy figure of embodiment
Ultraviolet visible spectrometry is the conventional means for studying Secondary structure, the Soret absorption bands of protein
Position migration can provide its structural information.Fig. 2 is HRP solution (curve a) and the HRP-AuNTs mixed liquors (UV- of curve b)
Vis abosrption spectrograms, Soret absorption bands are both present at 403.00nm, and wavelength location does not change substantially, show HRP with
Original conformation is still maintained after AuNTs mixing, is changed without recurring structure.
Embodiment 3
(1) 1.6g graphite powders, 0.8g ionic liquids HPPF are taken6It is placed in mortar with 500 μ L atoleines and grinds 3h, obtained
Then carbon paste is inserted in the vitreous electricity pole pipe that internal diameter is 4mm and is compacted by carbon paste, interpolation copper wire obtains CILE as conducting wire；
(2) the AuNTs solution drop coating that 10 μ L optical density (OD)s are 1 is taken to be protected from light naturally dry under room temperature on CILE surfaces,
Obtain AuNTs/CILE electrodes；
(3) 8 μ L 15mgmL are taken again-1HRP solution drop coating is protected from light nature in AuNTs/CILE electrode surfaces at room temperature
It dries, obtains HRP/AuNTs/CILE electrodes；
(4) 6 μ L 0.5%Nafion ethanol solutions drop coatings is finally taken to be kept away at room temperature in HRP/AuNTs/CILE electrode surfaces
Light dry after to get to Nafion/HRP/AuNTs/CILE electrodes.
Embodiment 4
(1) 2.5g graphite powders, 1.0g ionic liquids HPPF are taken6It is placed in mortar with 500 μ L atoleines and grinds 5h, obtained
Then carbon paste is inserted in the vitreous electricity pole pipe that internal diameter is 4mm and is compacted by carbon paste, interpolation copper wire obtains CILE as conducting wire；
(2) the AuNTs solution drop coating that 12 μ L optical density (OD)s are 1 is taken to be protected from light naturally dry under room temperature on CILE surfaces,
Obtain AuNTs/CILE electrodes；
(3) 9 μ L 20mgmL are taken again-1HRP solution drop coating is protected from light nature in AuNTs/CILE electrode surfaces at room temperature
It dries, obtains HRP/AuNTs/CILE electrodes；
(4) 8 μ L 0.7%Nafion ethanol solutions drop coatings is finally taken to be kept away at room temperature in HRP/AuNTs/CILE electrode surfaces
Light dry after to get to Nafion/HRP/AuNTs/CILE electrodes.
Direct Electrochemistries of 5 HRP of embodiment on modified electrode
Fig. 3 is cyclic voltammogram of the different modifying electrode in the PBS solution of pH 3.0.It can be seen from the figure that
(curve a) and Nafion/AuNTs/CILE (do not have oxidation peak reduction peak, show electrode surface not Nafion/CILE on curve b)
There are electroactive materials.(occur a pair of asymmetrical redox peaks on curve c) in Nafion/HRP/CILE, illustrate HRP
There are electronics transfer, still electron transfer rate is slower between CILE.And in Nafion/HRP/AuNTs/CILE (on curve d)
There are a pair of good and will definitely be inverse redox peaks to occur.The result shows that adding in AuNTs in electrode surface causes HRP
The redox peaks that electrochemical reaction generates become more symmetrical, and redox peak current is consequently increased, this is because in electricity
The AuNTs of high conductivity can improve the load capacity of HRP and accelerate the transfer of the electronics between basal electrode and HRP in pole surface,
Current-responsive is caused to increase.It is respectively Epa=-0.159V and Epc=-0.232V that spike potential can be immediately arrived at from curve d
(vs.SCE), spike potential poor (△ Ep) is 0.073V, Middle Eocene E0The ratio between '=- 0.196V (vs.SCE), redox peak current
Close to 1, the characteristic electrochemical of prosthetic heme group Fe (III) in redox protein matter/Fe (II) oxidation-reduction pair is shown
Scholarship and moral conduct is.
6 electrochemical AC impedance collection of illustrative plates of embodiment
Electrochemical alternate impedance spectrum (EIS) can effectively provide the impedance variations information of electrode face finish process, and
Electronics transfer resistance (Ret) can be read by the diameter of semicircle.Fig. 4 be respectively AuNTs/CILE (curve a),
Nafion/HRP/AuNTs/CILE (curve b), Nafion/CILE (curve c), the Nafion/HRP/CILE (exchanges of curve d)
Impedance spectra.Half diameter of a circle is equal to the size of impedance value, represents the resistance sizes of electrode surface.AuNTs/CILE (curves
A) for straight line almost without resistance, this is because the good conductivity of AuNTs.And Nafion/CILE (curve c) and
(impedance value of curve d) increases to 76.4 Ω and 217.6 Ω to Nafion/HRP/CILE, this is because Nafion and HRP are not
Conduction hinders [Fe (CN) in the presence of electrode surface6]3-/4-Electronics transfer, and then increase resistance value.And Nafion/
(impedance value of curve b) is 33.9 Ω to HRP/AuNTs/CILE, and reductions of impedance value illustrates that AuNTs has high electric conductivity and greatly
Specific surface area, reduce interface resistance, accelerate the transfer of electronics.
7 gold nano triangular plate of embodiment and horseradish peroxidase modified electrode restore the electro-catalysis of TCA
Using Nafion/HRP/AuNTs/CILE electro-catalysis reduction TCA, the results are shown in Figure 5.Delay in the PBS of pH 3.0
It rushes after solution adds in TCA and records cyclic voltammetry curve, find reduction peak occur in -0.254V, reduction peak current is dense with TCA
The increase of degree and increase, oxidation peak is reduced with the increase of TCA concentration even to disappear, and electro catalytic mechanism is as follows：
HRP Fe(III)+e→HRP Fe(II) (1)
2HRP Fe(II)+Cl3CCOOH+H+→2HRP Fe(III)+Cl2CHCOOH+Cl (2)
HRP Fe(II)+e→HRP Fe(I) (3)
2HRP Fe(I)+Cl2CHCOOH+H+→2HRP Fe(II)+ClCH2COOH+Cl (4)
2HRP Fe(I)+ClCH2COOH+H+→2HRP Fe(II)+CH3COOH+Cl (5)
When the concentration of TCA is when in the range of 0.1~250.0mmol/L, the concentration Cheng Liang of reduction peak current and TCA
Good linear relationship, equation of linear regression are I (μ A)=2.734C (mmolL-1)+28.91 (n=28, γ=0.996), inspection
Survey is limited to 0.033mmolL-1(3σ).When the concentration of TCA is more than 250.0mmolL-1When peak current occur a platform, electricity
Flow valuve remains unchanged, this is the characteristic feature of Michaelis-Menten dynamic processes.According to Lineweaver-Burk side
Journey：
Apparent K_m (K can be obtainedM app), it is that investigation enzyme-substrate reactions are dynamic (dynamical) heavy in enzymic catalytic reaction
Want index.I in formulassBe plus substrate after formed good linear relationship when steady-state current, C be add in substrate concentration, ImaxBe
The concentration of TCA is more than 250.0mmolL-1When the saturation of substrates state that is formed under measured maximum current.According to double works reciprocal
Figure method (1/Iss~1/ [TCA]) calculate the K that Nafion/HRP/AuNTs/CILE reacts the catalysis of TCAM appFor
176.4mmol·L-1, show that Nafion/HRP/AuNTs/CILE is good to the catalytic effect of TCA.This is because the height of AuNTs
Electric conductivity and bigger serface so that the electron transfer rate between HRP and modified electrode is strengthened, so as to enhance to TCA's
Catalytic effect.
6 gold nano triangular plate of embodiment and horseradish peroxidase modified electrode are to NaNO2Electro-catalysis reduction
Nafion/HRP/AuNTs/CILE electro-catalysis restores NaNO2The results are shown in Figure 6.Delay in the PBS of pH 3.0
It rushes solution and adds in NaNO2After record cyclic voltammetry curve, there is new reduction peak in -0.674V, reduction peak current is with NaNO2
The increase of concentration and increase.Work as NaNO2Concentration in 1.6~31.0mmolL-1In the range of when equation of linear regression be I
(μ A)=1.888C (mmolL-1) -0.0143 (n=31, γ=0.993), it detects and is limited to 0.53mmolL-1.It calculates
Nafion/HRP/AuNTs/CILE is to NaNO2It is catalyzed the K of reactionM appFor 27.77mmolL-1。
Claims (9)
1. one kind is based on gold nano triangular plate and horseradish peroxidase modified electrode, it is characterised in that the electrode includes substrate electricity
Pole, gold nano triangular plate (AuNTs) coating, horseradish peroxidase (HRP) coating, Nafion coatings, the basal electrode are
Ionic Liquid Modified carbon paste electrode (CILE).
2. a kind of electrode, it is characterised in that be followed successively by CILE basal electrodes, AuNTs coatings, HRP coatings, Nafion from inside to outside
The electrode of coating, i.e. Nafion/HRP/AuNTs/CILE electrodes.
3. a kind of electrochemica biological sensor part, it is characterised in that using claim 1-2 any one of them electrode as work
Electrode.
4. application of the claim 1-2 any one of them electrode in electrochemica biological sensor part is prepared.
5. the preparation method of claim 1-2 any one of them electrodes, it is characterised in that include the following steps：
(1) appropriate AuNTs solution drop coating is taken to be protected from light naturally dry under room temperature, obtain on CILE basal electrodes surface
AuNTs/CILE electrodes；
(2) appropriate HRP solution drop coating is taken to be protected from light naturally dry at room temperature, obtain HRP/ in AuNTs/CILE electrode surfaces
AuNTs/CILE electrodes；
(3) take appropriate Nafion ethanol solutions drop coating in HRP/AuNTs/CILE electrode surfaces, be protected from light at room temperature after drying to get
To Nafion/HRP/AuNTs/CILE electrodes.
6. the method described in claim 5, it is characterised in that the optical density (OD) of AuNTs solution described in step (1) is 1；Step
(2) a concentration of 10~20mgmL of the HRP solution-1, preferably 15mgmL-1, step (3) the Nafion ethanol solutions
Volumetric concentration for 0.3~0.7%, preferably 0.5%.
7. claim 5-6 any one of them methods, it is characterised in that the preparation method of step (1) the CILE basal electrodes
Include the following steps：By suitable graphite powder, N- hexyl pyridine hexafluorophosphates HPPF6, atoleine is placed in mortar and grinds
Uniformly, carbon paste is obtained, then carbon paste is inserted in vitreous electricity pole pipe and is compacted, interpolation copper wire obtains ionic liquid and repair as conducting wire
Adorn carbon paste electrode, i.e. CILE basal electrodes；Wherein, graphite powder and HPPF6Mass ratio be 1.5~2.5:1, preferably 2:1；Every gram
HPPF6Use 500~700 μ L atoleines, preferably 500 μ L；Milling time is 3.0-5.0h；Glass electrode bore is 4mm.
8. application of the claim 1-2 any one of them electrode in catalysis reduction trichloroacetic acid and/or sodium nitrite.
9. application of the claim 1-2 any one of them electrode in detection trichloroacetic acid and/or sodium nitrite.
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| CN114559029A (en) * | 2022-03-02 | 2022-05-31 | 广州大学 | Gold nanoparticles, preparation method and application thereof |
| CN115184430A (en) * | 2022-07-08 | 2022-10-14 | 天津大学 | Electrooxidation stress marker 8-hydroxyl deoxyguanosine detection electrode and device |
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| CN110672688A (en) * | 2019-08-30 | 2020-01-10 | 佛山市南海区里水镇经济促进局 | Electrochemical biosensor for detecting tetrabromobisphenol A bis (2-hydroxyethyl) ether and preparation method and application thereof |
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| CN114559029A (en) * | 2022-03-02 | 2022-05-31 | 广州大学 | Gold nanoparticles, preparation method and application thereof |
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| CN115184430A (en) * | 2022-07-08 | 2022-10-14 | 天津大学 | Electrooxidation stress marker 8-hydroxyl deoxyguanosine detection electrode and device |
| CN115184430B (en) * | 2022-07-08 | 2024-05-31 | 天津大学 | Electrode and device for detecting 8-hydroxydeoxyguanosine serving as electric oxidation stress marker |
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