CN104407027B - Method for preparing glutathione imprinted sensor through magnetic-control induced self-assembly and quick film formation - Google Patents
Method for preparing glutathione imprinted sensor through magnetic-control induced self-assembly and quick film formation Download PDFInfo
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Abstract
The invention discloses a method for preparing a glutathione imprinted sensor through magnetic-control induced self-assembly and quick film formation. The method sequentially comprises six steps as follows: electrode pretreatment, preparation of Fe3O4@PANI-rGO composite nanoparticles, joint pre-assembly of the composite nanoparticles and an imprinted layer compound, electrode modification through magnetic-control induced self-assembly, imprinted polymer formation through electric polymerization and template molecule elution. The preparation method is simple and convenient to operate, the price is low, the response is sensitive, the interference rejection is high, stability and repeatability are good, glutathione in blood plasma can be detected efficiently, flexibly and rapidly, and the method has important significance on diagnosis and treatment for clinical related diseases.
Description
Technical field
The invention belongs to technical field of analysis and detection is and in particular to a kind of magnetic control induction self assembly fast filming is used for preparing
The method of glutathione trace sensor.
Background technology
Glutathione (glutathione, GSH) is to be present in biological internal a kind of important non-protein sulfydryl chemical combination
Thing, plays an important role in vivo as a kind of important biological reductant, such as detoxication, maintenance cytotostatic
Property, anti-oxidant/nitration damage etc.[1].The elimination of GSH is closely related with the multiple disease of human body, such as diabetes, AIDS and god
Through DD etc.[2].Therefore, the content measuring GSH in biological sample has important meaning to the diagnosis of relevant disease and treatment
Justice and practical value.
Electrochemical sensor sensitivity is high, design is simple, cheap, have good stability and reusing, can
Realize real-time monitoring thus be widely used[3].But poor selectivity become its development bottleneck, and molecular imprinting have single-minded
Property strong, feature that anti-adverse environment ability is strong, the two is combined together and brings out the best in each other, in Food Inspection, environmental monitoring, chemistry
Have great application prospect in product and medicine production[4].The sensitive material that molecularly imprinted polymer is used as sensor has become point
One important application of sub- engram technology, calls this electrochemical sensor using molecularly imprinted polymer as sensitive material
Molecular imprinting electrochemical sensor.Its compared with the biological sensitive materials electrochemical sensor that Recent study is more, be difficult by
Biodegradable destruction, repeats and utilizes, and preparation is simple, and high temperature resistant, high pressure, acid, alkali and organic solvent, is therefore expected to become
The ideal substitute of biomaterial.
Molecular self-assembling has and can spontaneously form in situ, Thermodynamically stable, and the high defect of coverage is few, molecular ordered arrangement,
Can artificially design vector surface texture, the principal character such as simple and easy to get, prepare electrochemical element using MSA and become and grind now
The focus studied carefully.And the understanding being orientated behavior under magnetic field to particle with people progressively moves to maturity, magnetic field has become as one kind
New self assembly power.Self-assembling technique is induced to form the orderly new material of structure height using magnetic control, in magnetic field magnetization
So that the easy magnetizing axis of magnetic-particle unanimously arranges along magnetic direction under power effect, form one-dimensional orderly magnetic Nano knot
Structure.By induced by magnetic field self-assembling technique introduce MIPs be prepared as formed structurally ordered, the controlled polymer film of thickness creates
Advantage.
Magnetic nano particle has that biocompatibility is excellent, electric conductivity is high and the little and segregative advantage of toxic and side effect.Graphite
Alkene be a kind of hexagonal lattice constitute have perfect two dimensional crystal structure, sheet, individual layer, have excellent conduction,
Heat conduction and mechanical property, huge specific surface area, the development for novel nano electrochemical sensor provides good carrier.Polyphenyl
Amine because having various structure, unique mechanism of doping effect, excellent physical and chemical performance, good stability and raw material inexpensive
The advantages of be easy to get, and become the focus of polymer research.The combine tri compound nanoparticle of formation of three is made full use of three
The advantage planting nano material, obtains high-specific surface area, high conductivity, has the nano material of catalysis activity.At present, magnetic control is lured
The self assembly leading tri compound nanoparticle have not been reported for the preparation of glutathione molecules trace electrochemical sensor.
Content of the invention
This method adopts the study frontier magnetic control of self-assembling technique to induce self-assembling technique, is received with induced by magnetic field tri compound
Grain of rice self-assembled modified electrode.Under the influence of a magnetic field, Fe3O4@PANI-rGO(Fe3O4@polyaniline-reduced graphene) ternary
Composite nano-granule is assembled into structurally ordered network structure along outer magnetic field direction.Adjust magnetic field intensity and may be implemented in molecular level
The upper thickness controlling film, composition and structure, not only greatly strengthen the sensitivity of electrode after its modification, also orderly for MIPs film
The formation of structure creates advantage.On this modified electrode surface with glutathione as template molecule, Fe3O4@PANI-rGO is
Function monomer, pyrroles is common function monomer and crosslinking agent, and constant potential polymerization is prepared has efficient identification to glutathione
Highly sensitive molecular imprinting electrochemical sensor, and achieve the sensitive quick detection of plasma sample GSH-PX activity.The method
Fast and simple, there is good recovery and reappearance, the monitoring for clinical glutathione provides a kind of new method.
The purpose of the present invention is achieved in the following ways：
A kind of magnetic control induces self assembly fast filming to be used for the method preparing glutathione trace sensor, and the method includes
Following steps：
A) electrode pretreatment：Magnetic glassy carbon electrode is through A12O3After suspension polishing, absolute ethyl alcohol, deionized water is used to surpass respectively
Sound cleans；
b)Fe3O4Prepared by@PANI-rGO composite nanoparticle：By graphene oxide and Fe3O4@PANI composite nano-granule is pressed
It is 1 according to mass ratio：10～1：30 ratio is jointly pre-assembled, is subsequently adding hydrazine hydrate and is reduced, and constant temperature stirs 6～10h, magnetic
Property separate, wash to neutrality, obtain Fe3O4@PANI-rGO tri- primitive composite nano-granule；
C) composite nanoparticle is jointly pre-assembled with imprinted layer bond complexes：Prepare and contain function monomer, potassium chloride, sulphur
Acid, glutathione and Fe3O4The mixed solution of@PANI-rGO composite nano-granule, seals after nitrogen charging, places 2 in room temperature light protected environment
～8h, completes function monomer, glutathione and Fe3O4@PANI-rGO nanoparticle pre-assembled；
D) through induced by magnetic field self-assembled modified electrode：Take containing function monomer, glutathione and Fe3O4@PANI-rGO is multiple
Close the pre-assembled liquid of nanoparticle, after nitrogen blows, insertion magnetic pole absorption, magnetic field intensity is 0.32～1.6T；
E) electropolymerization forms imprinted polymer：Connect step d, with constant potential method electropolymerization, after taking-up, deionized water is repeatedly
Drip washing, nitrogen dries up；
F) eluted template molecule：With constant potential method in H2SO4In solution, electrochemistry wash-out, then anti-with a large amount of distilled waters
Multiple flushing, removes glutathione.
Step " c) " GSH-PX activity is 1 with the molar concentration rate of function monomer：30～1：50, function monomer is preferably five
Membered heterocycles, preferably five-membered heterocycles are pyrroles.
In step b), hydrazine hydrate act as reducing agent.
Insertion magnetic pole adsorption time is 5～20min in step " d) ", the magnetic field intensity of magnetic pole is 0.32～
The magnetic field intensity of 1.6T, preferably magnetic pole is 0.64～1.28T, most preferably 0.96T, and the process of this absorption is for magnetic control induction certainly
The process of assembling.
In constant potential method in step " e) ", constant potential is 0.3～0.8V；Constant potential polymerization time is 200～500s.
In step " f) ", the time of electroelution is 2～8min.
Preferably glutathione, Fe3O4@PANI-rGO nanoparticle, function monomer, the mass ratio of potassium chloride and sulfuric acid is 1:
0.26:4.4:6.2:0.03～1:0.46:13.1:12.4:0.39.
Fe3O4The preparation method of@PANI composite nano-granule is as follows：Containing Fe3O4In the hydrochloric acid solution of nanoparticle and aniline, its
Middle Fe3O4The mass ratio of nanoparticle and aniline is 1:1～1:3, add ammonium persulfate to react to obtain blackish green liquid at 0～5 DEG C,
Magnetic Isolation, washs to neutrality, is vacuum dried, obtains Fe3O4@PANI composite nano-granule.Containing Fe3O4Nanoparticle and the hydrochloric acid of aniline
In solution, the concentration of hydrochloric acid can be 0.1mol L-1.
Fe3O4The preparation of@PANI specifically can be according to following steps：Weigh 4～5g FeCl3 .6H2O and 1～3g
FeCl2 .4H2O in 250mL three-necked bottle, stirring and dissolving, temperature rises to and adds 8～10mL ammoniacal liquor react 1～3h when 80 DEG C.Pure
It is washed to neutrality, prepared Fe3O4Nanoparticle.The Fe that will be obtained3O4Nanoparticle is poured in 250mL beaker, and addition 100mL concentration is
0.1mol L-1Hydrochloric acid, add 1～6mL aniline.After dispersed with stirring half an hour, under ultrasound condition, dropping 8～10mL concentration is
0.1mol L-1Ammonium persulfate (per half a minute one), and control temperature to react 3h at 0～5 DEG C, obtain blackish green liquid.Magnetic
Property separate, wash to neutrality, be vacuum dried 24h, obtain Fe3O4@PANI composite nano-granule.
Fe3O4The preparation of@PANI-rGO：In the three-necked bottle of 100mL, take 6.5mg mL-1Graphene oxide (GO) 2～
8mL, is dissolved in 40mL deionized water, ultrasonic 1h.It is subsequently adding the Fe preparing3O4@PANI composite nano-granule, itself and graphite oxide
Alkene mass ratio is 30:1～10:1,80 DEG C of pre-assembled reaction 1h, add 0.2～0.4mL reducing agent hydrazine hydrate, constant temperature stirring 6～
12h.Magnetic Isolation, washs to neutrality, is vacuum dried 24h, obtains Fe3O4@PANI-rGO tri- primitive composite nano-granule.
2. by the following method electrochemical sensor of the present invention is detected, electrochemical sensor preparation process is with enforcement
Example 2：
Electrochemical detection method and condition：
Cyclic voltammetry (CV) method：CV method：Detection potential range is -0.2～0.6V, and sweep speed is 100mV s-1.
Differential pulse method (DPV) method：Detection potential range is -0.8～0.6V, and current potential increment is 0.004V, and amplitude is
0.06V, pulse width is 0.2s, and sampling width is 0.04s, and quiescent time is 4s.
Probe solution：L containing 1mmol-1The 0.1mol L of the potassium ferricyanide-1KCl solution.
Before test, electrode is in H2SO4In solution, electroelution recovers to background current.For making absorption completely, may be selected in experiment
The enrichment time of glutathione is 10min.
3. the glutathione molecules blotting membrane electrochemical sensor adopting magnetic control induction of the present invention is to concentration known solution
Static Adsorption is tested：The MIES preparing is taking concentration range respectively 2.0 × 10-8～1.0 × 10-6mol·L-1Gluathione
Peptide solution is tested, and measured concentration is respectively 2.0 × 10-8～5.0 × 10-5mol·L-1Become preferably linear, calculating can
Know that detection is limited to 3nmol L-1(S/N=3).
4. test sample process：Take infection from hospital experimenter's blood plasma, after pretreatment, prepared the PBS solution of pH 7.0, adopt
The concentration of DPV method determination sample GSH-PX activity.
5. the impact to blotting membrane for the change of magnetic field intensity：Investigate magnetic field intensity when 0.32T～1.6T changes, preparation
The response of trace sensor current change, find magnetic field intensity in 0.96T, current-responsive value is maximum, prepare under the conditions of this
Trace transducer sensitivity highest.Thickness and the surface knot of the blotting membrane being formed during change of magnetic field strength are further investigated
The change of structure.When magnetic field intensity is too low, during especially less than 0.64T, the trace film thickness being formed is less, although surface is uniformly
But sparse, now electric conductivity is poor and imprinting efficiency is low, and transducer sensitivity is poor.When magnetic field intensity is too high, during higher than 1.28T,
The trace film thickness being formed is larger, surface aggregation and uneven, the now too fast gathering of nanoparticle and template molecule embedded
Too deep lead to imprinting efficiency still relatively low, the sensitivity of sensor is still poor.When magnetic field intensity is 0.64～1.28T, institute
Moderate about 4.5～5.5 μm of the trace film thickness being formed, surface uniformly, is conducive to electrical conduction and improves imprinting efficiency, now
The sensitivity of trace sensor is higher.Compare the surface texture of the film being not added with magnetic place forming simultaneously, find to be not added with magnetic field institute
The film being formed is sparse and uneven, is further characterized by the effective power that magnetic field is the induction orderly self assembly of composite nano-granule, Fig. 6 energy
Spectrum phenogram further illustrates this result.
Beneficial effects of the present invention compared with the prior art：
1. molecular engram is combined with electrochemical sensor, improves the selectivity of electrochemical sensor it is achieved that blood plasma
The selective quick detection of sample GSH-PX activity.
2. adopt magnetic control self-assembling technique, be can achieve by the magnetic field intensity adjusting magnetic pole and control on a molecular scale
The thickness of film, composition and structure, not only greatly strengthen the sensitivity of electrode, shorten the self assembly time after its modification, and
Preparation is convenient, simple to operate, cheap, is also that the formation of MIPs film ordered structure creates advantage simultaneously.
3. have benefited from high conductivity and the catalysis activity of tri compound nanoparticle, amplify electrochemical signals, preparation further
High-sensitive glutathione trace sensor.
4. adopt the polypyrrole film layer of electropolymerization preparation stable, complete, fine and close, good conductivity, the combination situation with matrix
Preferably, considerably increase specific surface area and the sensitivity of electrode, the highly sensitive molecular engram of tool can be prepared using this property
Polymer film.
Brief description
Fig. 1 is the EIS phenogram (after d. absorption after c. wash-out before a. bare electrode b. wash-out) of MIES preparation process, shows magnetic
Field induction Fe3O4@PANI-rGO composite nano-granule be adsorbed onto electrode electropolymerization after (curve b), electrode impedance is with respect to bare electrode
(curve a) reduces, and Fe is described3O4The MIPs film conductivity that the doping of@PANI-rGO composite nano-granule is formed is good.Divide through template
(after curve c), electrode impedance reduces again it was demonstrated that template molecule is eluted and forms corresponding trace hole for son wash-out.And template
After Molecular Adsorption (curve d), electrode impedance increase be returned nearly to elute before level it was demonstrated that template molecule enters trace
Hole hinders potassium ferricyanide probe molecule to reach electrode surface.
Fig. 2 is the CV figure (after d. absorption after c. wash-out before a. bare electrode b. wash-out) of MIES preparation process.Induced by magnetic field
Fe3O4In the orderly self assembly of MGCE electrode surface, after electropolymerization, (curve b) forms the MIPs of dense uniform to@PANI-rGO nanoparticle
Film, with respect to bare electrode, (curve a) significantly increases peak point current.This is due to Fe3O4@PANI-rGO nano composite material is led
Superior electrical property, increased the current-responsive of electrode.After eluted template molecule, (curve c) peak point current increases further, shows
The imprinted polymer being embedded with template molecule before wash-out hinders probe molecule and is transferred to electrode surface, and after eluted template molecule
Imprinted polymer in define the peculiar identification hole of GSH, probe molecule can be made to pass through hole-transfer and to occur to electrode surface
Electrochemical redox reaction is thus produce the response current of amplification.After absorption template molecule, (curve d), peak point current is almost
Return to the level before wash-out and hinder probe molecule to reach electrode surface it was demonstrated that template molecule enters trace hole.
Fig. 3 has or not magnetic nano particle preparation MIES electrochemical response comparison diagram, and (a. bare electrode b. has nanoparticle c. no nanometer
Grain).Doping Fe3O4(curve b) is with respect to bare electrode (curve a) and no nanoparticle (curve c) for the composite nano-granule of@PANI-rGO
Peak point current significantly increases, and shows the Fe that adulterates3O4@PANI-rGO nanoparticle greatly increases the electric conductivity of MIES and electro-catalysis is lived
Property.
Fig. 4 tests current-responsive result figure for selective absorption.Common interference thing in other 5 kinds of human bodies is selected in this experiment
(ascorbic acid, uric acid, dopamine, cysteine, oxidized form of glutathione) has investigated the preparation of the embodiment of the present invention 1 method
MIES and the selection performance of non-molecular engram electrochemical sensor (NIES)：Electrode is measured in glutathione solution with DPV method
Current-responsive after absorption, records peak point current I0；Measure electrode with method respectively molten with the mixing of chaff interference containing glutathione
Peak point current I after adsorbing in liquidm, result show 20 times of concentration chaff interferences on MIES measure glutathione current-responsive affect very
Little, Im/I0Value be 0.92～1.1, show prepare sensor selectivity excellent.
Fig. 5 is the current differential figure before and after MIES wash-out during change of magnetic field strength.It can be seen that respond highest, △ in 0.96T
The value of Ip is 92 μ A.
Fig. 6 A-G is scanning electron microscope (SEM) photograph, and H is the energy spectrum diagram of the blotting membrane being formed when having or not magnetic field.
Wherein, A and D for magnetic field intensity for 0.32T when the blotting membrane that formed, B and E for magnetic field intensity for 0.96T when formed
Blotting membrane, C and F for magnetic field intensity for 1.6T when the blotting membrane that formed, G is to be not added with the surface texture that magnetic field forms film.
A-C shows the thickness of blotting membrane being formed during change of magnetic field strength, and figure A, B, C show, magnetic field intensity is respectively
When 0.32,0.96,1.6T, the thickness of the blotting membrane being formed is respectively 3.2 μm, 5.1 μm and 6.9 μm.D-F shows magnetic field intensity
The change of the trace membrane superficial tissue being formed during change.Figure D-F shows, with the change of magnetic field intensity, the surface texture of film is sent out
Give birth to change, illustrated to control microstructure and the thickness of formed blotting membrane by changing magnetic field intensity.Figure G is not
Plus the membrane superficial tissue of magnetic place forming, with respect to the uniform sequential structure being formed under induced by magnetic field, the structure on its surface is dilute
Dredge and uneven, show that magnetic field is the effective power of the induction orderly self assembly of tri compound nanoparticle.In figure H, I is magnetic field intensity
Energy spectrum diagram under 0.96T, II is the energy spectrum diagram of no magnetic field intensity, further demonstrate that the above results.
Specific embodiment
By the following examples explanation is further explained to the present invention：
Medicine and reagent：Al2O3(0.05 μm, Shanghai Chen Hua Instrument Ltd.), glutathione (reduced form) (try by West Asia
Agent research center), potassium chloride (KCl analyzes pure, chemical reagent Co., Ltd of group of nations), the potassium ferricyanide (K3Fe(CN)6, point
Analysis is pure, Shanghai newly precious Fine Chemical Works), absolute ethyl alcohol (analyzes pure, chemical reagent Co., Ltd of group of nations), six chloride hydrate
Iron (Chemical Reagent Co., Ltd., Sinopharm Group), Iron dichloride tetrahydrate (chemistry materials factory of Wenzhou City), ammonium persulfate (insult by Shanghai
Peak chemical reagent Co., Ltd), NaOH (NaOH, Nanjing Chemistry Reagent Co., Ltd.), hydrochloric acid (HCl, Liyang east chemistry
Reagent Co., Ltd), pyrroles's (pyrrole analyzes pure, Aladdin reagent Co., Ltd), sulfuric acid (H2SO4, Solution on Chemical Reagents in Shanghai
Co., Ltd), sodium dihydrogen phosphate (NaH2PO4, analyze pure, Nanjing Chemistry Reagent Co., Ltd.), disodium hydrogen phosphate (Na2HPO4,
Analyze pure, Shanghai Ling Feng Chemical Co., Ltd.), hydrazine hydrate (H2N-NH2, Shanghai Experimental Reagent Co., Ltd.), nitrogen (technical grade,
Nanjing 55 institute), experimental water is redistilled water.
In following examples, Fe3O4The preparation of@PANI：Weigh 4.72g FeCl3 .6H2O and 1.72g FeCl2 .4H2O in
In 250ml three-necked bottle, stirring and dissolving, temperature rises to and adds 10ml ammoniacal liquor when 80 DEG C and react 1h.Pure water is washed till neutrality, is obtained
Fe3O4Nanoparticle.The Fe that will be obtained3O4Nanoparticle is poured in 250mL beaker, and addition 100mL concentration is 0.1mol L-1Hydrochloric acid,
Add 4mL aniline.After dispersed with stirring half an hour, under ultrasound condition, dropping 8mL concentration is 0.1mol L-1Ammonium persulfate (every half
Minute one), and control temperature to react 3h at 0～5 DEG C, obtain blackish green liquid.Magnetic Isolation, washs to neutrality, vacuum is done
Dry 24h, obtains Fe3O4@PANI composite nano-granule.
Embodiment 1
A) electrode pretreatment：Magnetic glassy carbon electrode (Φ=5mm) is through 0.05 μm of A12O3After suspension polishing, use respectively no
Water-ethanol, deionized water are cleaned by ultrasonic 2min.
b)Fe3O4Prepared by@PANI-rGO composite nanoparticle：In the three-necked bottle of 100mL, take 6.5mg mL-1Oxidation stone
Black alkene (GO) 3mL, is dissolved in 40mL deionized water, ultrasonic 1h.It is subsequently adding the 0.5g Fe of preparation3O4@PANI composite nano-granule, 80
DEG C pre-assembled reaction 1h, adds 0.3mL reducing agent hydrazine hydrate, and constant temperature stirs 10h.Magnetic Isolation, washs to neutrality, is vacuum dried
24h, obtains Fe3O4@PANI-rGO tri- primitive composite nano-granule.
C) composite nanoparticle is jointly pre-assembled with imprinted layer bond complexes：Prepare L containing 2.5mmol-1GSH (gluathione
Peptide), 0.3mg L-1Fe3O4@PANI-rGO composite nano-granule, 100mmol L-1Pyrroles, 2mmol L-1H2SO4, 0.1mol L- 1The mixed aqueous solution 5mL of KCl, seals after nitrogen charging, stirs 5h, complete function monomer, template molecule and friendship in room temperature light protected environment
Join the pre-assembled of agent.
D) through induced by magnetic field self-assembled modified electrode：Take containing function monomer, glutathione and Fe3O4Pre- group of@PANI
Dress liquid, after nitrogen blows 10min, insertion magnetic pole absorption 5min, the magnetic field intensity of magnetic pole is 0.32T.
E) electropolymerization forms imprinted polymer：Connect step d, with constant potential method under 0.5V electropolymerization 400s, after taking-up,
Deionized water drip washing repeatedly, nitrogen dries up.
F) eluted template molecule：With constant potential method in H2SO4Electrochemistry wash-out 5min in solution, uses double steamings in a large number then
Water rinses repeatedly, removes template molecule.
Embodiment 2
A) electrode pretreatment：Magnetic glassy carbon electrode (Φ=5mm) is through 0.05 μm of A12O3After suspension polishing, use respectively no
Water-ethanol, deionized water are cleaned by ultrasonic 2min.
b)Fe3O4Prepared by@PANI-rGO composite nanoparticle：In the three-necked bottle of 100mL, 6.5mg mL-1Graphite oxide
Alkene (GO) 6mL, is dissolved in 40mL deionized water, ultrasonic 1h.It is subsequently adding the 0.5g Fe of preparation3O4@PANI composite nano-granule, 80 DEG C
Pre-assembled reaction 1h, adds 0.2mL reducing agent hydrazine hydrate, and constant temperature stirs 10h.Magnetic Isolation, washs to neutrality, is vacuum dried
24h, obtains Fe3O4@PANI-rGO tri- primitive composite nano-granule.
C) composite nanoparticle is jointly pre-assembled with imprinted layer bond complexes：Prepare L containing 2.5mmol-1GSH (gluathione
Peptide), 0.3mg L-1Fe3O4@PANI-rGO composite nano-granule, 75mmol L-1Pyrroles, 1mmol L-1H2SO4, 0.1mol L-1KCl
Mixed aqueous solution 5mL, after nitrogen charging seal, in room temperature light protected environment stir 6h, complete function monomer, template molecule and crosslinking agent
Pre-assembled.
D) through induced by magnetic field self-assembled modified electrode：Take containing function monomer, glutathione and Fe3O4Pre- group of@PANI
Dress liquid, after nitrogen blows 10min, insertion magnetic pole absorption 10min, the magnetic field intensity of magnetic pole is 0.96T.
E) electropolymerization forms imprinted polymer：Connect step d, with constant potential method under 0.5V electropolymerization 300s, after taking-up,
Deionized water drip washing repeatedly, nitrogen dries up.
F) eluted template molecule：With constant potential method in H2SO4Electrochemistry wash-out 4min in solution, uses double steamings in a large number then
Water rinses repeatedly, removes template molecule.
Embodiment 3
A) electrode pretreatment：Magnetic glassy carbon electrode (Φ=5mm) is through 0.05 μm of A12O3After suspension polishing, use respectively no
Water-ethanol, deionized water are cleaned by ultrasonic 2min.
b)Fe3O4Prepared by@PANI-rGO composite nanoparticle：In the three-necked bottle of 100mL, take 6.5mg mL-1Oxidation stone
Black alkene (GO) 5mL, is dissolved in 40mL deionized water, ultrasonic 1h.It is subsequently adding the 0.5g Fe of preparation3O4@PANI composite nano-granule, 80
DEG C pre-assembled reaction 1h, adds 0.3mL reducing agent hydrazine hydrate, and constant temperature stirs 10h.Magnetic Isolation, washs to neutrality, is vacuum dried
24h, obtains Fe3O4@PANI-rGO tri- primitive composite nano-granule.
C) composite nanoparticle is jointly pre-assembled with imprinted layer bond complexes：Prepare L containing 2.5mmol-1GSH (gluathione
Peptide), 0.3mg L-1Fe3O4@PANI-rGO composite nano-granule, 125mmol L-1Pyrroles, 1mmol L-1H2SO4, 0.1mol L- 1The mixed aqueous solution 5mL of KCl, seals after nitrogen charging, stirs 4h, complete function monomer, template molecule and friendship in room temperature light protected environment
Join the pre-assembled of agent.
D) through induced by magnetic field self-assembled modified electrode：Take containing function monomer, glutathione and Fe3O4Pre- group of@PANI
Dress liquid, after nitrogen blows 10min, insertion magnetic pole absorption 15min, the magnetic field intensity of magnetic pole is 1.28T.
E) electropolymerization forms imprinted polymer：Connect step d, with constant potential method under 0.6V electropolymerization 450s, after taking-up,
Deionized water drip washing repeatedly, nitrogen dries up.
F) eluted template molecule：With constant potential method in H2SO4Electrochemistry wash-out 6min in solution, uses double steamings in a large number then
Water rinses repeatedly, removes template molecule.
Embodiment 4
A) electrode pretreatment：Magnetic glassy carbon electrode (Φ=5mm) is through 0.05 μm of A12O3After suspension polishing, use respectively no
Water-ethanol, deionized water are cleaned by ultrasonic 2min.
b)Fe3O4Prepared by@PANI-rGO composite nanoparticle：In the three-necked bottle of 100mL, take 6.5mg mL-1Oxidation stone
Black alkene (GO) 4mL, is dissolved in 40mL deionized water, ultrasonic 1h.It is subsequently adding the 0.5g Fe of preparation3O4@PANI composite nano-granule, 80
DEG C pre-assembled reaction 1h, adds 0.2mL reducing agent hydrazine hydrate, and constant temperature stirs 10h.Magnetic Isolation, washs to neutrality, is vacuum dried
24h, obtains Fe3O4@PANI-rGO tri- primitive composite nano-granule.
C) composite nanoparticle is jointly pre-assembled with imprinted layer bond complexes：Prepare L containing 2.5mmol-1GSH (gluathione
Peptide), 0.3mg L-1Fe3O4@PANI-rGO composite nano-granule, 90mmol L-1Pyrroles, 2mmol L-1H2SO4, 0.1mol L-1KCl
Mixed aqueous solution 5mL, after nitrogen charging seal, in room temperature light protected environment stir 4h, complete function monomer, template molecule and crosslinking agent
Pre-assembled.
D) through induced by magnetic field self-assembled modified electrode：Take containing function monomer, glutathione and Fe3O4Pre- group of@PANI
Dress liquid, after nitrogen blows 10min, insertion magnetic pole absorption 10min, the magnetic field intensity of magnetic pole is 0.64T.
E) electropolymerization forms imprinted polymer：Connect step d, with constant potential method under 0.8V electropolymerization 350s, after taking-up,
Deionized water drip washing repeatedly, nitrogen dries up.
F) eluted template molecule：With constant potential method in H2SO4Electrochemistry wash-out 4min in solution, uses double steamings in a large number then
Water rinses repeatedly, removes template molecule.
Bibliography
[1]Gunaratnam M,Grant MH.Cr(VI)inhibits DNA,RNA and protein syntheses
in hepatocytes:Involvement of glutathione reductase,reduced glutathione and
DT-diaphorase[J].Toxicol.in Vitro,2008,22:879-886.
[2]Raffa M,Mechri A,Othman LB,et al.Decreased glutathione levels and
antioxidant enzyme activities in untreated and treated schizophrenic patients
[J].Prog.Neuro-Psychoph,2009,33:1178-1183.
[3]Zhu LL,Cao YH,Cao GQ.Electrochemical sensor based on magnetic
molecularly imprinted nanoparticles at surfactant modified magnetic electrode
for determination of bisphenol[J].Biosens.Bioelectron,2014,54:258-261.
[4]Li HF,Xie CG,Li SQ,Xu K.Electropolymerized molecular imprinting on
gold nanoparticle-carbon nanotube modified electrode for electrochemical
detection of triazophos[J].Colloid Surface B,2012,89:175-181.
Claims (7)
1. a kind of magnetic control induces self assembly fast filming to be used for preparing the method for glutathione trace sensor it is characterised in that being somebody's turn to do
Method comprises the following steps：
A) electrode pretreatment：Magnetic glassy carbon electrode is through A12O3After suspension polishing, use absolute ethyl alcohol, deionized water ultrasonic respectively
Cleaning；
b) Fe3O4Prepared by@PANI-rGO composite nanoparticle：By graphene oxide and Fe3O4@PANI composite nano-granule presses quality
Than for 1：10~1：30 ratio is jointly pre-assembled, is subsequently adding hydrazine hydrate and is reduced, constant temperature stirring 6 ~ 10 h, Magnetic Isolation,
Wash to neutrality, obtain Fe3O4@PANI-rGO tri- primitive composite nano-granule；
C) composite nanoparticle is jointly pre-assembled with imprinted layer bond complexes：Prepare and contain function monomer, potassium chloride, sulfuric acid, paddy
The sweet peptide of Guang and Fe3O4The mixed solution of@PANI-rGO composite nano-granule, seals after nitrogen charging, places 2 ~ 8 h in room temperature light protected environment,
Complete function monomer, glutathione and Fe3O4@PANI-rGO nanoparticle pre-assembled；Described function monomer is pyrroles；
D) through induced by magnetic field self-assembled modified electrode：Take containing function monomer, glutathione and Fe3O4@PANI-rGO is combined
The pre-assembled liquid of nanoparticle, after nitrogen blows, insertion magnetic pole absorption, magnetic field intensity is 0.32 ~ 1.6 T；
E) electropolymerization forms imprinted polymer：Connect step d, with constant potential method electropolymerization, after taking-up, deionized water is drenched repeatedly
Wash, nitrogen dries up；Constant potential is 0.3 ~ 0.8 V；
F) eluted template molecule：With constant potential method in H2SO4In solution, electrochemistry wash-out, is rushed repeatedly with a large amount of distilled waters then
Wash, remove glutathione.
2. magnetic control according to claim 1 induces self assembly fast filming to be used for preparing the side of glutathione trace sensor
Method is it is characterised in that its GSH-PX activity is 1 with the molar concentration rate of function monomer：30~1：50.
3. the preparation method of molecular engram film electrochemical sensor according to claim 1 is it is characterised in that step " d) "
Middle insertion magnetic pole adsorption time is 5 ~ 20 min.
4. the preparation method of molecular engram film electrochemical sensor according to claim 1 is it is characterised in that step " e) "
Middle constant potential is 0.3 ~ 0.8 V.
5. the preparation method of molecular engram film electrochemical sensor according to claim 1 is it is characterised in that step " e) "
Middle constant potential polymerization time is 200 ~ 500 s.
6. the preparation method of molecular engram film electrochemical sensor according to claim 1 is it is characterised in that step " f) "
The time of middle electroelution is 2 ~ 8 min.
7. the preparation method of molecular engram film electrochemical sensor according to claim 1 is it is characterised in that Fe3O4@PANI
The preparation method of composite nano-granule is as follows：Containing Fe3O4In the hydrochloric acid solution of nanoparticle and aniline, wherein Fe3O4Nanoparticle and aniline
Mass ratio be 1: 1~1 :3, add ammonium persulfate in 0～5oReact to obtain blackish green liquid, Magnetic Isolation under C, wash to
Neutrality, vacuum drying, obtain Fe3O4@PANI composite nano-granule.
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| CN104910411B (en) * | 2015-06-18 | 2018-04-20 | 辽宁师范大学 | The method that electric field-assisted removing template molecule prepares protein-imprinted polymer |
| CN105891295B (en) * | 2016-03-29 | 2018-06-05 | 南京医科大学 | Based on electric conductivity trace nanoparticle and magnetic control induction filming to the electrochemical detection method of insulin |
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