CN104034778A - Chitosan-ionic liquid-graphene-enzyme composite membrane modified electrode and preparation method thereof - Google Patents
Chitosan-ionic liquid-graphene-enzyme composite membrane modified electrode and preparation method thereof Download PDFInfo
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- CN104034778A CN104034778A CN201410272483.0A CN201410272483A CN104034778A CN 104034778 A CN104034778 A CN 104034778A CN 201410272483 A CN201410272483 A CN 201410272483A CN 104034778 A CN104034778 A CN 104034778A
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Abstract
The invention discloses a chitosan-ionic liquid-graphene-enzyme composite membrane modified electrode and a preparation method of the chitosan-ionic liquid-graphene-enzyme composite membrane modified electrode. The preparation method comprises the following steps: (1) dissolving chitosan into a dilute acid solution to obtain a solution A; (2) adding graphene oxide into the solution A, and performing ultrasonic decomposition to obtain a solution B; (3) adding an ionic liquid into the solution B, and stirring to obtain a solution C; (4) adding enzyme into the solution C, and performing ultrasonic treatment to obtain a solution D; and (5) putting a working electrode into the solution D, and performing constant potential deposition, thereby obtaining the electrode. The electrode has the remarkable characteristics that firstly, the preparation process is simple and convenient, and the cost is low; secondly, the ratio and the thickness of the composite membrane can be controlled through a constant potential deposition method and reactant amount; and thirdly, the chitosan is rich in resource, good in biocompatibility and is an excellent enzyme immobilization carrier, and due to addition of the ionic liquid and the graphene, good electron transmission channels for connecting an oxidation reduction center of enzyme with the surface of the electrode are formed, and relatively good bioactivity of the enzyme is kept.
Description
Technical field
The invention belongs to the preparation field of modified electrode, be specifically related to the preparation method of a kind of shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode.
Background technology
Shitosan, as a kind of inexhaustible, nexhaustible renewable resource, is the product of chitin process part deacetylation, and it is the straight-chain polysaccharide coupling together by β-Isosorbide-5-Nitrae glycosidic bond, and its chemical constitution is with amino alkaline polysaccharide polymkeric substance.Shitosan has good good biocompatibility and unique bioactivation function, by successfully for the fixation support of enzyme.Ionic liquid is as a kind of green solvent, compare with electrolyte aqueous solution with traditional organic solvent, having the advantages such as non-volatile, good electric conductivity, wider electrochemical window and selective dissolution ability, is used widely in the fields such as synthetic at organic catalysis, galvanochemistry is synthetic, biological chemistry.In recent years, it is found that many enzymes have good performance in ionic liquid, be conducive to carry out galvanochemistry and catalytic reaction.Graphene is as desirable Two-dimensional Carbon based nano-material, due to its monoatomic layer thickness and two s' planar structure, for it provides great specific surface area and excellent electric conductivity.Therefore using Graphene and compound thereof as modified electrode, the research that builds electrochemica biological sensor has caused that people pay close attention to greatly.
Modified electrode is as the vitals in electrochemica biological sensor, and its preparation method has a great impact the performance of sensor.The controllability of enzyme modified electrode preparation process is fast for meeting with a response, highly sensitive, the sensor of good stability, long service life is most important.But, in conventional enzyme modified electrode preparation, the compound substance of preparing need to be dispersed in water again by chemical method at present, then drip and be coated onto electrode surface, drop-coating is consuming time more, the bad dispersibility of gained film, and coating is coarse, thickness is difficult to accurate control, causes the combination property of modified electrode to decline.
Summary of the invention
The object of the present invention is to provide a kind of shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode and preparation method thereof, the cathodic reduction of Graphene is combined with the electro-deposition electricity of shitosan, single stage method has obtained shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode, compare with at present conventional enzyme modified electrode preparation method, preparation condition is easy, and technique is simple.
The present invention solves the problems of the technologies described above adopted technical scheme: shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode, and the product that it obtains for following preparation method, includes following steps:
1) shitosan is dissolved in 0.01-1mol/L dilute acid soln, obtains solution A, in solution A, the mass percent concentration of shitosan is 0.5-5%;
2) in solution A, add 5-20mg graphene oxide, ultrasonic dispersion 1-2 hour obtains solution B;
3) in solution B, add ionic liquid, stir 1-2 hour, obtain solution C, ionic liquid quality is the 1-5% of solution B quality;
4) in solution C, add enzyme, ultrasonic 10-30 minute, obtains solution D, and the concentration of enzyme in solution D is 0.01-1mg/mL;
5) working electrode is placed in to solution D, carries out potentiostatic electrodeposition, obtain shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode.
Press such scheme, described diluted acid is hydrochloric acid or acetic acid.
Press such scheme, described ionic liquid is 1-butyl-3-methylimidazole phosphorus hexafluoride or 1-butyl-3-methylimidazole tetrafluoride boron.
Press such scheme, described working electrode is gold, platinum, glass carbon or indium tin oxide-coated glass.
Press such scheme, the current potential of described potentiostatic electrodeposition is-0.5～-2V; The time of described potentiostatic electrodeposition is 1-10 minute.
Press such scheme, described enzyme is peroxidase, Lactate Oxidase, lipase or cholesterol oxidase.
The present invention has following distinguishing feature: 1) cathodic reduction of Graphene is combined with the electro-deposition electricity of shitosan, single stage method obtains complex film modified electrode, and the preparation technology of complex film modified electrode is easy, and cost is low; 2) composite membrane proportioning and thickness can control by potentiostatic electrodeposition method and reactant consumption; 3) shitosan source is abundant, having good biocompatibility, is good enzyme immobilization carrier, and the adding of ionic liquid and Graphene, make the redox center of ligase and the good electron propagation ducts between electrode surface, and make enzyme keep good biologically active.
Embodiment
For a better understanding of the present invention, below in conjunction with embodiment, further illustrate content of the present invention, but content of the present invention is not only confined to the following examples.
Embodiment 1:
1) shitosan is dissolved in 0.1mol/L dilute hydrochloric acid solution, obtains solution A, in solution A, the mass percent concentration of shitosan is 0.5%;
2) in solution A, add 5mg graphene oxide, ultrasonic dispersion obtains solution B for 1 hour;
3) in solution B, add 1-butyl-3-methylimidazole phosphorus hexafluoride, stir 1 hour, obtain solution C, ionic liquid quality is 2% of solution B quality;
4) in solution C, add peroxidase, ultrasonic 10 minutes, obtain solution D, the concentration of enzyme in solution D is 0.05mg/mL;
5) indium tin oxide-coated glass electrode is placed in to solution D, carries out potentiostatic electrodeposition, sedimentation potential is-0.7V, and sedimentation time is 2 minutes, obtains shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode,
6) shitosan-ionic liquid-Graphene obtaining-enzyme composite membrane modified electrode for concentration of hydrogen peroxide 3.2 * 10
-4-1.5 * 10
-3within the scope of mol/L, there is good inspection effect.
Embodiment 2:
1) shitosan is dissolved in 0.5mol/L dilute acetic acid solution, obtains solution A, in solution A, the mass percent concentration of shitosan is 1%;
2) in solution A, add 10mg graphene oxide, ultrasonic dispersion obtains solution B for 1 hour;
3) in solution B, add 1-butyl-3-methylimidazole phosphorus hexafluoride, stir 2 hours, obtain solution C, ionic liquid quality is 1% of solution B quality;
4) in solution C, add lipase, ultrasonic 20 minutes, obtain solution D, the concentration of enzyme in solution D is 0.1mg/mL;
5) gold electrode is placed in to solution D, carries out potentiostatic electrodeposition, sedimentation potential is-1V, and sedimentation time is 5 minutes, obtains shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode,
6) shitosan-ionic liquid-Graphene obtaining-enzyme composite membrane modified electrode for glyceryl dioleate concentration 3.0 * 10
-6-2 * 10
-4within the scope of mg/mL, there is good inspection effect.
Embodiment 3:
1) shitosan is dissolved in 0.5mol/L dilute hydrochloric acid solution, obtains solution A, in solution A, the mass percent concentration of shitosan is 2%;
2) in solution A, add 10mg graphene oxide, ultrasonic dispersion obtains solution B for 2 hours;
3) in solution B, be incorporated as 1-butyl-3-methylimidazole phosphorus hexafluoride, stir 1 hour, obtain solution C, ionic liquid quality is 3% of solution B quality;
4) in solution C, add cholesterol oxidase, ultrasonic 30 minutes, obtain solution D, the concentration of enzyme in solution D is 0.1mg/mL;
5) platinum electrode is placed in to solution D, carries out potentiostatic electrodeposition, sedimentation potential is-1.2V, and sedimentation time is 4 minutes, obtains shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode,
6) shitosan-ionic liquid-Graphene obtaining-enzyme composite membrane modified electrode for cholesterol concentration 2.0 * 10
-5-3 * 10
-4within the scope of mol/L, there is good inspection effect.
Embodiment 4:
1) shitosan is dissolved in 1mol/L dilute acetic acid solution, obtains solution A, in solution A, the mass percent concentration of shitosan is 1%;
2) in solution A, add 5mg graphene oxide, ultrasonic dispersion obtains solution B for 1 hour;
3) in solution B, add 1-butyl-3-methylimidazole tetrafluoride boron, stir 1 hour, obtain solution C, ionic liquid quality is 5% of solution B quality;
4) in solution C, add Lactate Oxidase, ultrasonic 15 minutes, obtain solution D, the concentration of enzyme in solution D is 0.3mg/mL;
5) glass-carbon electrode is placed in to solution D, carries out potentiostatic electrodeposition, sedimentation potential is-1.1V, and sedimentation time is 6 minutes, obtains shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode,
6) shitosan-ionic liquid-Graphene obtaining-enzyme composite membrane modified electrode for lactic acid concn 1.0 * 10
-4-9 * 10
-3within the scope of mol/L, there is good inspection effect.
Embodiment 5:
1) shitosan is dissolved in 0.7mol/L dilute hydrochloric acid solution, obtains solution A, in solution A, the mass percent concentration of shitosan is 3%;
2) in solution A, add 15mg graphene oxide, ultrasonic dispersion obtains solution B for 2 hours;
3) in solution B, add 1-butyl-3-methylimidazole tetrafluoride boron, stir 1 hour, obtain solution C, ionic liquid quality is 3% of solution B quality;
4) in solution C, add lipase, ultrasonic 10 minutes, obtain solution D, the concentration of enzyme in solution D is 0.2mg/mL;
5) glass-carbon electrode is placed in to solution D, carries out potentiostatic electrodeposition, sedimentation potential is-1.6V, and sedimentation time is 2 minutes, obtains shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode,
6) shitosan-ionic liquid-Graphene obtaining-enzyme composite membrane modified electrode for glyceryl dioleate concentration 2.0 * 10
-6-1.0 * 10
-4within the scope of mg/mL, there is good inspection effect.
Embodiment 6:
1) shitosan is dissolved in 0.8mol/L dilute acetic acid solution, obtains solution A, in solution A, the mass percent concentration of shitosan is 1%;
2) in solution A, add 10mg graphene oxide, ultrasonic dispersion obtains solution B for 1 hour;
3) in solution B, add 1-butyl-3-methylimidazole tetrafluoride boron, stir 2 hours, obtain solution C, ionic liquid quality is 5% of solution B quality;
4) in solution C, add peroxidase, ultrasonic 15 minutes, obtain solution D, the concentration of enzyme in solution D is 0.3mg/mL;
5) gold electrode is placed in to solution D, carries out potentiostatic electrodeposition, sedimentation potential is-0.8V, and sedimentation time is 8 minutes, obtains shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode,
6) shitosan-ionic liquid-Graphene obtaining-enzyme composite membrane modified electrode for concentration of hydrogen peroxide 2.0 * 10
-4-1.0 * 10
-3within the scope of mol/L, there is good inspection effect.
Each cited raw material of the present invention can be realized the present invention, and the bound value of each raw material, interval value can realize the present invention; At this, do not enumerate embodiment.Bound value, the interval value of technological parameter of the present invention can be realized the present invention, at this, do not enumerate embodiment.
Claims (12)
1. shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode, the product that it obtains for following preparation method, includes following steps:
1) shitosan is dissolved in 0.01-1mol/L dilute acid soln, obtains solution A, in solution A, the mass percent concentration of shitosan is 0.5-5%;
2) in solution A, add 5-20mg graphene oxide, ultrasonic dispersion 1-2 hour obtains solution B;
3) in solution B, add ionic liquid, stir 1-2 hour, obtain solution C, ionic liquid quality is the 1-5% of solution B quality;
4) in solution C, add enzyme, ultrasonic 10-30 minute, obtains solution D, and the concentration of enzyme in solution D is 0.01-1mg/mL;
5) working electrode is placed in to solution D, carries out potentiostatic electrodeposition, obtain shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode.
2. shitosan-ionic liquid-Graphene according to claim 1-enzyme composite membrane modified electrode, is characterized in that: described diluted acid is hydrochloric acid or acetic acid.
3. shitosan-ionic liquid-Graphene according to claim 1-enzyme composite membrane modified electrode, is characterized in that: described ionic liquid is 1-butyl-3-methylimidazole phosphorus hexafluoride or 1-butyl-3-methylimidazole tetrafluoride boron.
4. shitosan-ionic liquid-Graphene according to claim 1-enzyme composite membrane modified electrode, is characterized in that: described working electrode is gold, platinum, glass carbon or indium tin oxide-coated glass.
5. shitosan-ionic liquid-Graphene according to claim 1-enzyme composite membrane modified electrode, is characterized in that: the current potential of described potentiostatic electrodeposition is-0.5～-2V; The time of described potentiostatic electrodeposition is 1-10 minute.
6. shitosan-ionic liquid-Graphene according to claim 1-enzyme composite membrane modified electrode, is characterized in that: described enzyme is peroxidase, Lactate Oxidase, lipase or cholesterol oxidase.
7. the preparation method of shitosan-ionic liquid-Graphene claimed in claim 1-enzyme composite membrane modified electrode, includes following steps:
1) shitosan is dissolved in 0.01-1mol/L dilute acid soln, obtains solution A, in solution A, the mass percent concentration of shitosan is 0.5-5%;
2) in solution A, add 5-20mg graphene oxide, ultrasonic dispersion 1-2 hour obtains solution B;
3) in solution B, add ionic liquid, stir 1-2 hour, obtain solution C, ionic liquid quality is the 1-5% of solution B quality;
4) in solution C, add enzyme, ultrasonic 10-30 minute, obtains solution D, and the concentration of enzyme in solution D is 0.01-1mg/mL;
5) working electrode is placed in to solution D, carries out potentiostatic electrodeposition, obtain shitosan-ionic liquid-Graphene-enzyme composite membrane modified electrode.
8. the preparation method of shitosan-ionic liquid-Graphene according to claim 7-enzyme composite membrane modified electrode, is characterized in that: described diluted acid is hydrochloric acid or acetic acid.
9. the preparation method of shitosan-ionic liquid-Graphene according to claim 7-enzyme composite membrane modified electrode, is characterized in that: described ionic liquid is 1-butyl-3-methylimidazole phosphorus hexafluoride or 1-butyl-3-methylimidazole tetrafluoride boron.
10. the preparation method of shitosan-ionic liquid-Graphene according to claim 7-enzyme composite membrane modified electrode, is characterized in that: described working electrode is gold, platinum, glass carbon or indium tin oxide-coated glass.
The preparation method of 11. shitosan-ionic liquid-Graphene according to claim 7-enzyme composite membrane modified electrodes, is characterized in that: the current potential of described potentiostatic electrodeposition is-0.5～-2V; The time of described potentiostatic electrodeposition is 1-10 minute.
The preparation method of 12. shitosan-ionic liquid-Graphene according to claim 7-enzyme composite membrane modified electrodes, is characterized in that: described enzyme is peroxidase, Lactate Oxidase, lipase or cholesterol oxidase.
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Application publication date: 20140910 |