CN101839884A - 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 modified electrode 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 self assembly layer by layer and preparation method thereof of a kind of inorganic nano sheet and two kinds of albumen particularly is provided.
Background technology
(the Layer-by-layer assembly of self-assembly method successively of electrostatic precipitation, LBL) ultimate principle is: be driving force with the electrostatic interaction, at substrate surface, the polyelectrolyte alternating deposit of oppositely charged structure has component and the accurate controlled multilayer film system of thickness.This technology has that equipment needed thereby is simple, and the operating process controllability is strong, operating conditions gentleness, characteristics such as applied widely.Because protein, enzyme, biomolecule such as DNA itself have electric charge or can induce electric charge under certain conditions and become can be assembled polyelectrolyte, electrostatic precipitation in recent years successively self-assembling method also is widely used in field of biosensors.
At present, based on the biology sensor of static self-assembling method (LBL) preparation layer by layer 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 specific substrates.Yet, along with development of science and technology and growth in the living standard, people in productive life as environment measuring, health care, aspects such as pharmaceutical preparation detect require more and more higher to complex system and complex material, the electrode that needs the existence of a kind of dual-active component of exploitation or multiple active components is to reach the collaborative or facilitation of function between each active component, and the function of integrated two kinds of components realizes detecting the variation of 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, with two kinds of protein and hydrotalcite nano piece alternately on blank electrode deposition obtain the controlled multi-layer ultrathin membrane of Stability Analysis of Structures component.
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 a 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 of this modified electrode is evenly continuous, and ultrathin membrane presents the stack manner of long-range order on 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 adds 1-3 times of volume then is the ammonia spirit of 7.5-8.5, obtains shelling the hydrotalcite nano piece colloidal solution of layer;
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 handle 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, repeating alternately, immersion process promptly obtains hydrotalcite nano piece and double protein ultrathin film modified electrode with compounded up to reaching the required number of plies.
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: utilizing static self-assembling method layer by layer, is carrier with the inorganic material hydrotalcite nano piece, and two kinds of protein alternately are assembled into the ITO electrode surface uniformly, obtains Stability Analysis of Structures, the ultrathin film modified electrode of thickness homogeneous and controllable.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 stable.
Description of drawings
The different number of plies ultrathin membranes (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that Fig. 1 obtains for embodiment 1
nThe uv-visible absorption spectra spectrogram; Horizontal ordinate is a wavelength, unit: nanometer (nm), ordinate are absorbance, no 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 the absorbance of ultrathin membrane in 410 nanometers correspondences.
Fig. 2 obtains different number of plies ultrathin membranes (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) for embodiment 1
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 an intensity.
(hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that Fig. 3 obtains for embodiment 1
2The flat scanning electron microscope of ultrathin membrane and atomic force microscope figure.
(hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that Fig. 4 obtains for embodiment 1
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 the 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 a film thickness, and unit is nanometer (nm).
Fig. 5 is a 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 a voltage, unit: volt (V), with respect to the Ag/AgCl electrode, ordinate is an electric current, unit: microampere (μ A).
(hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase) that Fig. 6 obtains for embodiment 1
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 a voltage, unit: volt (V), and with respect to the Ag/AgCl electrode, ordinate is an 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 an 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, solution in there-necked flask 97 ℃ reflux down and stirred 24 hours, use 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
2Protection is strong agitation down, reacts 24 hours, takes 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 adds 1.5 times of volumes then is 8 ammonia spirit, obtains shelling the hydrotalcite nano piece colloidal solution of layer;
B. be 8.0 phosphate buffered solution dissolving haemoglobin with the pH value, hemoglobin concentration 1g/L, phosphate buffered solution 0.1M; With the pH value is 8.0 phosphate buffered solution dissolving horseradish peroxidase, 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; Be placed on 1.25g/L then, 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 1g/L again 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, promptly obtain (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
10Ultrathin film modified electrode.
Uv absorption by assembling ultrathin membrane on the electrode characterizes the growth for Thin Film process, behind assembling one deck horseradish peroxidase, carry out the test of a uv-visible absorption spectra, the ultraviolet-visible spectrophotometric 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 Fig. 2 respectively, 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 orderly arrangement in hydrotalcite layers.As can be seen, the modified electrode surface that the present invention constructs is evenly continuous on the ultrathin membrane surface scan Electronic Speculum of Fig. 3 and the atomic force microscope figure, 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.Ultrathin membrane thickness with the different numbers of plies is mapped to its number of plies, calculates every layer of ultrathin 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, on Shanghai occasion China CHI660 electrochemical workstation, carry out, ultrathin film modified electrode with the present invention's preparation is a working electrode, platinum filament is to electrode, silver/silver chloride electrode is that contrast electrode is a three-electrode system, serves as to detect the chemical property that end liquid detects the electrode of constructing among the present invention with the phosphate buffered solution of 0.1M.Blank ITO electrode and (hydrotalcite nano piece/haemoglobin/hydrotalcite nano piece/horseradish peroxidase)
2The cyclic voltammetric test 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 belong to the characteristic peak for 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 voltammetric test 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 following 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 excellent 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 following 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 excellent 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, solution in there-necked flask 97 ℃ reflux down and stirred 24 hours, use the deionized water centrifuging, wash to the pH value less than 8,70 ℃ of dryings 24 hours, obtain CoAl-CO
3LDH is standby.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
2Protection is strong agitation down, reacts 24 hours, takes 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 adds 2 times of volumes then is 8 ammonia spirit, obtains shelling the hydrotalcite nano piece colloidal solution of layer;
B. be 8.0 phosphate buffered solution dissolving horseradish peroxidase with the pH value, horseradish peroxidase concentration 1g/L, phosphate buffered solution 0.1M; With the pH value is 8.0 phosphate buffered solution dissolving glucose oxidase, 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; Be placed on 1.25g/L then, 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 1g/L again 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 a working electrode, and platinum filament is to electrode, and silver/silver chloride electrode is that contrast electrode is formed electrochemica biological sensor.Test temperature is 25 ℃, is that 7.0 phosphate buffered solution serves as to detect end liquid with 0.1M pH value, detects the response of ultrathin film modified electrode to glucose with timing ampere method, and 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 following 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 excellent 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 6g (0.15mol) NaOH in addition 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 adds 1 times of volume then is 8 ammonia spirit, obtains shelling the hydrotalcite nano piece colloidal solution of layer;
B. be 8.0 phosphate buffered solution dissolving haemoglobin with the pH value, hemoglobin concentration 1g/L, phosphate buffered solution 0.1M; With the pH value is 8.0 phosphate buffered solution dissolving myoglobins, 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; Be placed on 1.25g/L then, 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 1g/L again 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 a working electrode, and platinum filament is to electrode, and silver/silver chloride electrode is that contrast electrode is formed electrochemica biological sensor.Test temperature is 25 ℃, is that 7.0 phosphate buffered solution serves as to detect end liquid with 0.1M pH value, 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 following air after, to the H of 10 μ M
2O
2Response current keep original 90%, illustrate that the electrode that this method prepares has excellent storage stability.
Claims (7)
1. hydrotalcite nano piece and double protein ultrathin film modified electrode with compounded, it is characterized in that, this modified electrode is that hydrotalcite nano piece and two kinds of protein molecule alternating deposits are on basic electrode 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 a 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 of this modified electrode is evenly continuous, and ultrathin membrane presents the stack manner of long-range order on the direction perpendicular to the basic electrode surface, and the thickness nanoscale of ultrathin membrane is controlled.
2. the preparation method of hydrotalcite nano piece according to claim 1 and double protein ultrathin film modified electrode with compounded is characterized in that, its preparation process is as follows:
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 adds 1-3 times of volume then is the ammonia spirit of 7.5-8.5, obtains shelling the hydrotalcite nano piece colloidal solution of layer;
B. be 0.05-2M with concentration respectively with a-protein and PROTEIN B, the pH value is the phosphate buffered solution dissolving of 7-8, and the concentration of a-protein and PROTEIN B is respectively 0.5-2g/L;
C. will handle 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, repeating alternately, immersion process promptly obtains hydrotalcite nano piece and double protein ultrathin film modified electrode with compounded up to reaching the required number of plies.
3. according to the preparation method of right 2 described a kind of hydrotalcite nano pieces and double protein ultrathin film modified electrode with compounded, it is characterized in that 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+
4. according to the preparation method of right 2 described a kind of hydrotalcite nano pieces and double protein ultrathin film modified electrode with compounded, it is characterized in that described hydrotalcite precursor adopts coprecipitation, nucleation crystallization/isolation method, non-equilibrium crystallization method, urea method, ion exchange process or hydrothermal synthesis method preparation.
5. according to the preparation method of right 2 described a kind of hydrotalcite nano pieces and double protein ultrathin film modified electrode with compounded, it is characterized in that, 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.
6. according to the preparation method of right 2 described a kind of hydrotalcite nano pieces and double protein ultrathin film modified electrode with compounded, it is characterized in that, the described ITO electrode of step c disposal route is: the ITO electrode is respectively at deionized water, ethanol, acetone, methyl alcohol, deionized water for ultrasonic 5-30 minute, behind the deionized water rinsing, nitrogen dries up.
7. according to the preparation method of right 2 described a kind of hydrotalcite nano pieces and double protein ultrathin film modified electrode with compounded, it is characterized in that, step c described in electronegative macromolecule polymer solution immersion process be in the polyetherimide of 8-9.5 5-30 minute 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 the kayexalate of 0.5-2g/L and soaked 5-30 minute, dry up with the deionized water rinsing electrode surface and with nitrogen.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103616422A (en) * | 2013-12-03 | 2014-03-05 | 江南大学 | Preparation method of graphene/hemoglobin composite film modified electrode |
| CN106198653A (en) * | 2016-06-26 | 2016-12-07 | 北京化工大学 | A kind of sensor material of qualitative and quantitative detection glucose and preparation method thereof |
| 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 |
| CN109689881A (en) * | 2016-09-16 | 2019-04-26 | 伊士曼化工公司 | The biological sensor electrode prepared by physical vapour deposition (PVD) |
| US11630075B2 (en) | 2016-09-16 | 2023-04-18 | Eastman Chemical Company | Biosensor electrodes prepared by physical vapor deposition |
| US11835481B2 (en) | 2016-06-15 | 2023-12-05 | Eastman Chemical Company | Physical vapor deposited biosensor components |
| US11881549B2 (en) | 2017-06-22 | 2024-01-23 | Eastman Chemical Company | Physical vapor deposited electrode for electrochemical sensors |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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| Publication number | Priority date | Publication date | Assignee | Title |
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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 |
Non-Patent Citations (1)
| Title |
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| 《The Analyst》 20060221 Jun Wang等 Amperometric choline biosensor fabricated through electrostatic assembly of bienzyme/polyelectrolyte hybrid layers on carbon nanotubes 第479页第1段，第481页第18-24行 1-7 第131卷, 第4期 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103616422A (en) * | 2013-12-03 | 2014-03-05 | 江南大学 | Preparation method of graphene/hemoglobin composite film modified electrode |
| US11835481B2 (en) | 2016-06-15 | 2023-12-05 | Eastman Chemical Company | Physical vapor deposited biosensor components |
| CN106198653A (en) * | 2016-06-26 | 2016-12-07 | 北京化工大学 | A kind of sensor material of qualitative and quantitative detection glucose and preparation method thereof |
| 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 |
| CN109689881A (en) * | 2016-09-16 | 2019-04-26 | 伊士曼化工公司 | The biological sensor electrode prepared by physical vapour deposition (PVD) |
| US11624723B2 (en) | 2016-09-16 | 2023-04-11 | Eastman Chemical Company | Biosensor electrodes prepared by physical vapor deposition |
| US11630075B2 (en) | 2016-09-16 | 2023-04-18 | 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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| CN101839884B (en) | 2013-10-16 |
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