CN101839884B - Ultrathin film modified electrode with compounded hydrotalcite nanometer slices and double protein, and preparation method thereof - Google Patents
Ultrathin film modified electrode with compounded hydrotalcite nanometer slices and double protein, and preparation method thereof Download PDFInfo
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Abstract
The invention discloses an ultrathin film modified electrode constructed through the alternate layer-by-layer self-assembly of inorganic nanometer slices and two types of protein in the technical field of electrochemical biosensors, and a preparation method thereof. By an electrostatic precipitation layer-by-layer self-assembly method, the preparation method alternately assembles hydrotalcite nanometer slices and two different types of protein in hemoglobin, myoglobin, horseradish peroxidase and glucose oxidase onto the surface of an ITO electrode so as to construct an ultrathin film modifiedelectrode with compounded hydrotalcite nanometer slices/double protein. The ultrathin film constructed by the invention is uniform and continuous in surface, stable in structure and controllable in thickness and nanometer scale. The modified electrode has the advantages of efficient catalytic ability, good sensitivity, good operation stability and storage stability.
Description
Technical field
The invention belongs to the electrochemica biological sensor technical field, alternately ultrathin film modified electrode of constructing of layer by layer self assembly and preparation method thereof of a kind of inorganic nano sheet and two kinds of albumen particularly is provided.
Background technology
Successively self-assembly method (the Layer-by-layer assembly of electrostatic precipitation, LBL) ultimate principle is: take electrostatic interaction as driving force, at substrate surface, the polyelectrolyte alternating deposit of oppositely charged structure has component and the accurate controlled Multilayer system of thickness.This technology has the characteristics such as equipment needed thereby is simple, and the operating process controllability is strong, and operating conditions is gentle, and is applied widely.Because protein, enzyme, the biomolecule such as DNA itself with electric charge or can induce electric charge under certain conditions and become can be assembled polyelectrolyte, in recent years electrostatic precipitation successively self-assembling method also is widely used in field of biosensors.
At present, based on the biology sensor of layer by layer self-assembling method (LBL) preparation of static generally be with the chain complex polyelectrolyte particle or (with) inorganic nano material plants the biomolecule assembling with single, obtain the composite multilayer membrane modified electrode and realize detection performance to specific substrates.Yet, development and growth in the living standard along with science and technology, people in productive life such as environment measuring, health care, the aspects such as pharmaceutical preparation are more and more higher to complex system and complex material testing requirement, need the electrode of a kind of double activated component of exploitation or multiple active components existence to reach the collaborative or facilitation of function between each active component, the function of integrated two kinds of components realizes the variation of detection substrate and the optimization of sensitivity and interference free performance.
Summary of the invention
The object of the present invention is to provide a kind of hydrotalcite nano piece and double protein ultrathin film modified electrode with compounded and preparation method thereof.In conjunction with the rigid structure of inorganic nano sheet and static self-assembling method layer by layer, two kinds of protein and hydrotalcite nano piece are alternately obtained the controlled multi-layer ultrathin membrane of Stability Analysis of Structures component in blank electrode deposition.
The hydrotalcite nano piece of the present invention preparation and double protein ultrathin film modified electrode with compounded are that hydrotalcite nano piece and two kinds of protein molecule alternating deposits are on basic electrode tin indium oxide (ITO) electrode, it consists of the hydrotalcite nano lamella, the a-protein layer, the hydrotalcite nano lamella, the PROTEIN B layer repeats successively to assemble, and is labeled as (hydrotalcite nano piece/a-protein/hydrotalcite nano piece/PROTEIN B)
n, n is cycle index, a-protein and PROTEIN B are respectively any one in haemoglobin, myoglobins, horseradish peroxidase, the glucose oxidase, and a-protein and PROTEIN B are different proteins; The surface uniform of this modified electrode is continuous, and ultrathin membrane presents the stack manner of long-range order in the direction perpendicular to the basic electrode surface, and the thickness nanoscale of ultrathin membrane is controlled.
A. preparing interlayer anion is NO
3 -, laminate divalence, trivalent metal cation mol ratio M
2+/ M
3+The hydrotalcite precursor of=2.0-4.0; With 0.05-2g hydrotalcite precursor under nitrogen protection in 50-200mL formamide high speed stirring reaction 24-96 hour, the pH that then adds 1-3 times of volume is the ammonia spirit of 7.5-8.5, obtains the hydrotalcite nano piece colloidal solution of delamination;
B. with a-protein and PROTEIN B respectively with concentration for O.05-2M, the pH value is that the phosphate buffered solution of 7-8 is dissolved, the concentration of a-protein and PROTEIN B is respectively 0.5-2g/L;
C. will process clean ITO electrode soaks in electronegative macromolecule polymer solution and makes negative electricity on the electrode band, dry up with the deionized water rinsing electrode surface and with nitrogen, then it alternately is immersed in hydrotalcite nano piece colloidal solution 5-30 minute that step a obtains, the a-protein solution of step b preparation 5-30 minute, the hydrotalcite nano piece colloidal solution that step a obtains 5-30 minute, the PROTEIN B solution of step b preparation 5-30 minute, all use the deionized water rinsing electrode after soaking in the alternation procedure, and nitrogen dries up, repeat to replace immersion process until reach the required number of plies, namely obtain hydrotalcite nano piece and double protein ultrathin film modified electrode with compounded.
Described hydrotalcite laminate divalent metal is Mg
2+, Zn, Co
2+, Ni
2+, Ca
2+, Cu
2+, Fe
2+Or Mn
2+, trivalent metal cation is Al
3+, Cr
3+, Ga
3+, In
3+, Co
3+, Fe
3+Or V
3+
Described hydrotalcite precursor adopts coprecipitation, nucleation crystallization/isolation method, Non-Equilibrium Crystallization method, urea method, ion exchange process or hydrothermal synthesis method preparation.
Described a-protein and PROTEIN B are respectively any one in haemoglobin, myoglobins, horseradish peroxidase, the glucose oxidase, and a-protein and PROTEIN B are different proteins.
The described ITO electrode of step c disposal route is: at deionized water, ethanol, acetone, methyl alcohol, deionized water for ultrasonic 5-30 minute, behind the deionized water rinsing, nitrogen dries up the ITO electrode respectively.
Step c described in electronegative macromolecule polymer solution immersion process be middle 5-30 minute of the polyetherimide (PEI) of 8-9.5 for electrode being immersed in 0.5-2.5g/L, pH value, dry up with deionized water rinsing and with nitrogen, then electrode is immersed in middle the immersion 5-30 minute of kayexalate (PSS) of 0.5-2g/L, dries up with the deionized water rinsing electrode surface and with nitrogen.
The invention has the advantages that: utilize layer by layer self-assembling method of static, take the inorganic material hydrotalcite nano piece as carrier, two kinds of protein alternately are assembled into the ITO electrode surface uniformly, obtain Stability Analysis of Structures, the ultrathin film modified electrode that even thickness is controlled.Egg-pair white matter of the present invention and inorganic material ultrathin film modified electrode have shown good response signal, efficiently catalytic performance and good stability.
Description of drawings
Fig. 1 is the different number of plies ultrathin membranes (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that embodiment 1 obtains
nThe uv-visible absorption spectra spectrogram; Horizontal ordinate is wavelength, unit: nanometer (nm), and ordinate is absorbance, without unit; Illustration is mapped to the number of plies for the absorbance of each number of plies, and horizontal ordinate is the number of plies of ultrathin membrane, and ordinate is that ultrathin membrane is in absorbance corresponding to 410 nanometers.
Fig. 2 is that embodiment 1 obtains different number of plies ultrathin membranes (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
nXRD spectra, a wherein, b, the n value of c is respectively 3,8 and 15; Horizontal ordinate is 2 times of angles (2 θ), unit: degree; Ordinate is intensity.
Fig. 3 is (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that embodiment 1 obtains
2The flat scanning electron microscope of ultrathin membrane and atomic force microscope figure.
Fig. 4 is (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that embodiment 1 obtains
2The cross section scanning electron microscope diagram of ultrathin membrane; Figure a, b, c is respectively n=1, and 3,5; Figure d is the graph of a relation of different number of plies film thicknesses and the corresponding number of plies, and horizontal ordinate is the number of plies of ultrathin membrane, and ordinate is film thickness, and unit is nanometer (nm).
Fig. 5 is cyclic voltammetry curve, (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that a: embodiment 1 obtains
2Ultrathin film modified electrode, b: blank ITO electrode; Horizontal ordinate is voltage, unit: volt (V), with respect to the Ag/AgCl electrode, ordinate is electric current, unit: microampere (μ A).
Fig. 6 is (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that embodiment 1 obtains
2The circulation of the ultrathin film modified electrode curve of bending over the desk, solution is respectively (a) 10 μ mol/L, (b) 20 μ mol/L, (c) 30 μ mol/L, (d) 50 μ mol/L, (e) 80 μ mol/L, (f) 120 μ mol/L, (g) the adjacent phenol solution of 170 μ mol/L; Wherein horizontal ordinate is voltage, unit: volt (V), and with respect to the Ag/AgCl electrode, ordinate is electric current, unit: microampere (μ A); Illustration is the relation curve of catalytic current and adjacent phenol concentration, and wherein, horizontal ordinate is adjacent phenol concentration, unit: micromoles per liter (μ mol/L), ordinate is electric current, unit: microampere (μ A).
Embodiment
Embodiment 1:
A. the preparation of hydrotalcite nano piece colloidal solution: take by weighing 0.1mol Ni (NO
3)
26H
2O, 0.05molAl (NO
3) 39H
2O and 0.3mol urea are dissolved in the 500mL deionized water, and solution 97 ℃ of lower return stirrings 24 hours in there-necked flask are used the deionized water centrifuging, wash to the pH value near 7,70 ℃ of dryings 24 hours; Get 0.5g solid product and 63.75g NaNO
3Put into there-necked flask, add 500mL and take off CO
2Deionized water drips 0.16mL HNO again
3At N
2The lower strong agitation of protection was reacted 24 hours, took off CO
2The deionized water centrifuging is washed three times, and is used the ethanol suction filtration, and solid obtains the hydrotalcite precursor at 70 ℃ of vacuum drying 24h; Get 0.1g hydrotalcite precursor under nitrogen protection in the 100ml formamide strong agitation reaction 48h, the pH that then adds 1.5 times of volumes is 8 ammonia spirit, obtains the hydrotalcite nano piece colloidal solution of delamination;
B. be 8.0 phosphate buffered solution dissolving haemoglobin with the pH value, hemoglobin concentration 1g/L, phosphate buffered solution 0.1M; Be 8.0 phosphate buffered solution dissolving horseradish peroxidase with the pH value, horseradish peroxidase 1g/L, phosphate buffered solution 0.1M;
C.ITO electrode in deionized water, ethanol, acetone, methyl alcohol, deionized water each ultrasonic 10 minutes respectively, N
2Dry up; Then be placed on 1.25g/L, the pH value is to soak 20 minutes in 9 the electropositivity polyetherimide, behind deionized water rinsing, and N
2Dry up, the electronegative polyelectrolyte kayexalate solution of putting into again 1g/L soaked 20 minutes, and a large amount of negative charge on the electrode surface band dries up with the deionized water rinsing electrode surface and with nitrogen; Then it alternately is immersed in the horseradish peroxidase solution 25 minutes of the hemoglobin solutions 15 minutes of hydrotalcite nano piece colloidal solution 15 minutes that step a obtains, step b preparation, hydrotalcite nano piece colloidal solution 15 minutes that step a obtains, step b preparation, all use the deionized water rinsing electrode after soaking in the alternation procedure, and nitrogen dries up, repeat to replace immersion process 10 times, namely obtain (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
10Ultrathin film modified electrode.
Characterize the growth course of film by the uv absorption of assembling ultrathin membrane on the electrode, behind assembling one deck horseradish peroxidase, carry out the test of a uv-visible absorption spectra, the spectrophotometry test result of the ultrathin membrane of the different numbers of plies as shown in Figure 1, the 410nm place is the characteristic absorption band of haemoglobin and horseradish peroxidase activated centre protoheme porphyrin ring π-π * transition, the existence of this absorption band and increase increase by two kinds of protein of explanation being fixed in the substrate by the hydrotalcite nano piece success of absorbance along with cycle index.Illustration is the figure that 410nm place absorption value is done the number of plies of duplicature, and as can be seen from the figure, the protoheme porphyrin ring is in characteristic absorption peak even, linear increase along with the increase of the assembling number of plies at 410nm place.Protein successively being assembled in the substrate by success is described.Protein ultraviolet characteristic absorption maintenance be the best evidence that its conformation keeps, protein of this explanation assembling has well kept its original conformation in the laminate of hydrotalcite.
(hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) of the different numbers of plies of preparation
10The structure of ultrathin film modified electrode and surface topography XRD, SEM and AFM characterization result are seen respectively Fig. 2, Fig. 3 and Fig. 4.In the XRD spectra of Fig. 2, the superlattice diffraction peak that ° has occurred hydrotalcite in 2 θ=0.95, the d value is for being about 9.1nm, remove laminate thickness 0.96 nanometer of two hydrotalcites, the interlayer width is about 8.14nm, molecular dimension according to haemoglobin: 5.0 * 5.5 * 6.4nm, the molecular dimension of horseradish peroxidase: 3.0 * 3.5 * 6.0nm, can infer that haemoglobin and horseradish peroxidase arrange with unimolecular layer in hydrotalcite layers, increase along with cycle index n value, diffraction peak intensity is constantly strengthened, and illustrates that horseradish peroxidase is ordered arrangement in hydrotalcite layers.Can find out on the ultrathin membrane surface scan Electronic Speculum of Fig. 3 and atomic force microscope figure, the modified electrode surface that the present invention constructs is evenly continuous, and roughness is very little.The cross section of different number of plies ultrathin membranes is carried out the characterization result of scanning electron microscope and see Fig. 4, can find that from Fig. 4 the thickness of film increases uniformly along with the increase of the number of plies of assembling, illustrate that this method can realize that the nanoscale of film thickness is controlled.Membrane thickness with the different numbers of plies is mapped to its number of plies, calculates every layer of membrane thickness average out to 9.6nm from figure, and is similar to the thickness that XRD calculates.
Fig. 5 is (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
10The electrochemical test of ultrathin film modified electrode, carry out at Shanghai occasion China CHI660 electrochemical workstation, take the ultrathin film modified electrode of the present invention preparation as working electrode, platinum filament is to electrode, silver/silver chloride electrode is that contrast electrode is three-electrode system, detects the chemical property of the electrode of constructing among the present invention as detecting end liquid take the phosphate buffered solution of 0.1M.Blank ITO electrode and (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
2The cyclic voltammetry of ultrathin film modified electrode as shown in Figure 5, a pair of redox peak that will definitely be contrary has appearred in the prepared electrode of the present invention, and this can be attributed to the characteristic peak of haemoglobin and horseradish peroxidase activated centre protoheme prothetic group Fe (III)/Fe (II) generation redox reaction.Illustrate that the used process for fixation of the present invention can effectively promote the Direct Electrochemistry of protein.Same electrode scanning scans 10 times under the same conditions, and electric current is almost constant, illustrates that the modified electrode of the present invention's preparation has good operational stability.When modified electrode under 4 ℃, keep in the air after 2 months it being carried out multiple scanning, find that its peak current almost remains unchanged.Illustrate that protein fixation method has very high storage stability among the present invention.
With prepared (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
2The cyclic voltammetry result that ultrathin film modified electrode carries out catalysis to the catechol of variable concentrations as shown in Figure 6, increase along with catechol concentration in the end liquid, reduction peak constantly strengthens and oxidation peak remains unchanged, reduction peak current and catechol concentration relationship such as Fig. 6, as shown in Figure 6, the range of linearity of ultrathin film modified electrode catalysis catechol is 10-170 μ M, and related coefficient is 0.9992, and calculating this modified electrode by the straight line of Fig. 6 is 0.126 μ A/ μ M to the catalysis sensitivity of catechol.The prepared electrode pair catechol of this explanation has very strong sensitivity and catalytic performance.To 3 electrodes with a collection of preparation, the standard deviation of the response current of the catechol of its catalysis 10 μ M is 2.0%, shows (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
2The catalytic process of electrode pair glucose is reliable and stable.When modified electrode is preserved 2 months in 4 ℃ of lower air after, the response current of the catechol of 10 μ M is kept original 85%, illustrate that the electrode that this method prepares has good storage stability.To 3 electrodes with a collection of preparation, the standard deviation of the response current of the catechol of its catalysis 10 μ M is 2.0%, shows (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
2The catalytic process of electrode pair catechol is reliable and stable.When modified electrode is preserved 2 months in 4 ℃ of lower air after, the response current of the catechol of 10 μ M is kept original 88%, illustrate that the electrode that this method prepares has good storage stability.
Embodiment 2:
A. the preparation of hydrotalcite nano piece colloidal solution: claim 0.05mol Co (NO
3)
26H
2O, 0.025molAl (NO
3)
39H
2O and 0.175mol urea are dissolved in the 500mL deionized water, and solution 97 ℃ of lower return stirrings 24 hours in there-necked flask are used the deionized water centrifuging, wash to the pH value less than 8,70 ℃ of dryings 24 hours, obtain CoAl-CO
3LDH is for subsequent use.Get the CoAl-CO that 0.5g makes
3LDH and 63.75g NaNO
3Put into there-necked flask, add 500mL and take off CO
2Deionized water drips 0.16mL HNO again
3At N
2The lower strong agitation of protection was reacted 24 hours, took off CO
2The deionized water centrifuging is washed three times, and is used the ethanol suction filtration, and solid obtained CoAl-NO in 24 hours 70 ℃ of vacuum drying
3The hydrotalcite precursor; Get 0.1g hydrotalcite precursor under nitrogen protection in the 100ml formamide strong agitation reaction 48h, the pH that then adds 2 times of volumes is 8 ammonia spirit, obtains the hydrotalcite nano piece colloidal solution of delamination;
B. be 8.0 phosphate buffered solution dissolving horseradish peroxidase with the pH value, horseradish peroxidase concentration 1g/L, phosphate buffered solution 0.1M; Be 8.0 phosphate buffered solution dissolving glucose oxidase with the pH value, glucose oxidase 1g/L, phosphate buffered solution 0.1M;
C.ITO electrode in deionized water, ethanol, acetone, methyl alcohol, deionized water each ultrasonic 10 minutes respectively, N
2Dry up; Then be placed on 1.25g/L, the pH value is to soak 20 minutes in 9 the electropositivity polyetherimide, behind deionized water rinsing, and N
2Dry up, the electronegative polyelectrolyte kayexalate solution of putting into again 1g/L soaked 20 minutes, and a large amount of negative charge on the electrode surface band dries up with the deionized water rinsing electrode surface and with nitrogen; Then it alternately is immersed in the glucose oxidase solution 15 minutes of the horseradish peroxidase solution 15 minutes of hydrotalcite nano piece colloidal solution 15 minutes that step a obtains, step b preparation, hydrotalcite nano piece colloidal solution 15 minutes that step a obtains, step b preparation, all use the deionized water rinsing electrode after soaking in the alternation procedure, and nitrogen dries up, repeat to replace immersion process 2 times, can obtain (hydrotalcite nano piece/horseradish peroxidase/hydrotalcite nano piece/glucose oxidase)
2Ultrathin film modified electrode.
(hydrotalcite nano piece/horseradish peroxidase/hydrotalcite nano piece/glucose oxidase) with the present invention's preparation
2Ultrathin film modified electrode is working electrode, and platinum filament is to electrode, and silver/silver chloride electrode is that contrast electrode forms electrochemica biological sensor.Test temperature is 25 ℃, and take the phosphate buffered solution of 0.1M pH value as 7.0 as detecting end liquid, with the response of timing amperometric detection ultrathin film modified electrode to glucose, the range of linearity of this electrode detection glucose is 3.00 * 10
-5-2.4 * 10
-3Mol/L, related coefficient be 0.999. to 3 electrodes with a collection of preparation, the standard deviation of the response current of the glucose of its catalysis 10 μ M is 2.0%, shows (hydrotalcite nano piece/horseradish peroxidase/hydrotalcite nano piece/glucose oxidase)
2The catalytic process of electrode pair glucose is reliable and stable.When modified electrode is preserved 2 months in 4 ℃ of lower air after, the response current of the glucose of 10 μ M is kept original 85%, illustrate that the electrode that this method prepares has good storage stability.
Embodiment 3:
A. the preparation of hydrotalcite nano piece colloidal solution: take by weighing 12.82g (0.05mol) Mg (NO
3)
26H
2O and 6.27g (0.025mol) Al (NO
3)
39H
2O is dissolved in 250mL and takes off CO
2Be made into A solution in the deionized water, take by weighing in addition 6g (0.15mol) NaOH and be dissolved in 250mL and take off CO
2Be made into B solution in the deionized water.A solution and B solution are prepared MgAl-NO according to the method that nucleation/crystallization isolation method prepares hydrotalcite
3The hydrotalcite precursor, after the nucleation, crystallization was taken off CO after 15 hours under 100 ℃ of conditions
2The deionized water centrifuging is washed three times, and is used the ethanol suction filtration, and solid obtained MgAl-NO in 24 hours 70 ℃ of vacuum drying
3The hydrotalcite precursor, get 0.1g hydrotalcite precursor under nitrogen protection in the 100ml formamide strong agitation reaction 80h, the pH that then adds 1 times of volume is 8 ammonia spirit, obtains the hydrotalcite nano piece colloidal solution of delamination;
B. be 8.0 phosphate buffered solution dissolving haemoglobin with the pH value, hemoglobin concentration 1g/L, phosphate buffered solution 0.1M; Be 8.0 phosphate buffered solution dissolving myoglobins with the pH value, myoglobins 1g/L, phosphate buffered solution 0.1M;
C.ITO electrode in deionized water, ethanol, acetone, methyl alcohol, deionized water each ultrasonic 10 minutes respectively, N
2Dry up; Then be placed on 1.25g/L, the pH value is to soak 20 minutes in 9 the electropositivity polyetherimide, behind deionized water rinsing, and N
2Dry up, the electronegative polyelectrolyte kayexalate solution of putting into again 1g/L soaked 20 minutes, and a large amount of negative charge on the electrode surface band dries up with the deionized water rinsing electrode surface and with nitrogen; Then it alternately is immersed in the myoglobins solution 15 minutes of the hemoglobin solutions 15 minutes of hydrotalcite nano piece colloidal solution 15 minutes that step a obtains, step b preparation, hydrotalcite nano piece colloidal solution 15 minutes that step a obtains, step b preparation, all use the deionized water rinsing electrode after soaking in the alternation procedure, and nitrogen dries up, repeat to replace immersion process 2 times, can obtain (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/myoglobins)
2Ultrathin film modified electrode.
(hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/myoglobins) with the present invention's preparation
2Ultrathin film modified electrode is working electrode, and platinum filament is to electrode, and silver/silver chloride electrode is that contrast electrode forms electrochemica biological sensor.Test temperature is 25 ℃, take the phosphate buffered solution of 0.1M pH value as 7.0 as detecting end liquid, detects ultrathin film modified electrode to H with cyclic voltammetry
2O
2Response, this electrode detection H
2O
2The range of linearity be 2.00 * 10
-6-1 * 10
-4Mol/L, related coefficient be 0.999. to 3 electrodes with a collection of preparation, the H of its catalysis 10 μ M
2O
2The standard deviation of response current be 2.0%, show (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/myoglobins)
2Electrode pair H
2O
2Catalytic process reliable and stable.When modified electrode is preserved 2 months in 4 ℃ of lower air after, to the H of 10 μ M
2O
2Response current keep original 90%, illustrate that the electrode that this method prepares has good storage stability.
Claims (1)
1. the preparation method of a hydrotalcite nano piece and double protein ultrathin film modified electrode with compounded is characterized in that, its preparation process is as follows:
A. the preparation of hydrotalcite nano piece colloidal solution: take by weighing 0.1 mol Ni (NO
3)
26H
2O, 0.05 mol Al (NO
3)
39H
2O and 0.3mol urea are dissolved in the 500 mL deionized waters, and solution 97 ℃ of lower return stirrings 24 hours in there-necked flask are used the deionized water centrifuging, wash to the pH value near dry 24 hours of 7,70 oC; Get 0.5 g solid product and 63.75g NaNO
3Put into there-necked flask, add 500 mL and take off CO
2Deionized water drips 0.16mL HNO again
3At N
2The lower strong agitation of protection was reacted 24 hours, took off CO
2The deionized water centrifuging is washed three times, and is used the ethanol suction filtration, and solid obtains the hydrotalcite precursor at 70 ℃ of vacuum drying 24h; Get 0.1g hydrotalcite precursor under nitrogen protection in the 100ml formamide strong agitation reaction 48h, the pH that then adds 1.5 times of volumes is 8 ammonia spirit, obtains the hydrotalcite nano piece colloidal solution of delamination;
B. be 8.0 phosphate buffered solution dissolving haemoglobin with the pH value, hemoglobin concentration 1g/L, phosphate buffered solution 0.1M; Be 8.0 phosphate buffered solution dissolving horseradish peroxidase with the pH value, horseradish peroxidase 1g/L, phosphate buffered solution 0.1M;
C. the ITO electrode is in deionized water, ethanol, acetone, methyl alcohol, deionized water each ultrasonic 10 minutes respectively, N
2Dry up; Then be placed on 1.25g/L, the pH value is to soak 20 minutes in 9 the electropositivity polyetherimide, behind deionized water rinsing, and N
2Dry up, the electronegative polyelectrolyte kayexalate solution of putting into again 1g/L soaked 20 minutes, and a large amount of negative charge on the electrode surface band dries up with the deionized water rinsing electrode surface and with nitrogen; Then it alternately is immersed in the horseradish peroxidase solution 25 minutes of the hemoglobin solutions 15 minutes of hydrotalcite nano piece colloidal solution 15 minutes that step a obtains, step b preparation, hydrotalcite nano piece colloidal solution 15 minutes that step a obtains, step b preparation, all use the deionized water rinsing electrode after soaking in the alternation procedure, and nitrogen dries up, repeat to replace immersion process 10 times, namely obtain (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
10Ultrathin film modified electrode.
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| CN103616422A (en) * | 2013-12-03 | 2014-03-05 | 江南大学 | Preparation method of graphene/hemoglobin composite film modified electrode |
| CN109312384B (en) | 2016-06-15 | 2022-12-30 | 伊士曼化工公司 | Physical vapor deposition biosensor assembly |
| CN106198653B (en) * | 2016-06-26 | 2019-04-09 | 北京化工大学 | A kind of sensor material and preparation method thereof of qualitative and quantitative detection glucose |
| KR102547063B1 (en) | 2016-09-16 | 2023-06-22 | 이스트만 케미칼 컴파니 | Biosensor electrodes fabricated by physical vapor deposition |
| EP3512958B1 (en) | 2016-09-16 | 2022-04-06 | Eastman Chemical Company | Biosensor electrodes prepared by physical vapor deposition |
| US11881549B2 (en) | 2017-06-22 | 2024-01-23 | Eastman Chemical Company | Physical vapor deposited electrode for electrochemical sensors |
| CN109520813A (en) * | 2018-12-21 | 2019-03-26 | 马德君 | A kind of method of bivalent metal ion dissolution in detection hydrotalcite-based compound |
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| CN101393160A (en) * | 2008-10-29 | 2009-03-25 | 北京化工大学 | Biological functional multilayer film modified electrode and method for making same |
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