CN101196487B - Preparation method of modified electrode by electrodeposition of chitosan-ionic liquid-enzyme composite film - Google Patents
Preparation method of modified electrode by electrodeposition of chitosan-ionic liquid-enzyme composite film Download PDFInfo
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- CN101196487B CN101196487B CN2007101648420A CN200710164842A CN101196487B CN 101196487 B CN101196487 B CN 101196487B CN 2007101648420 A CN2007101648420 A CN 2007101648420A CN 200710164842 A CN200710164842 A CN 200710164842A CN 101196487 B CN101196487 B CN 101196487B
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Abstract
本发明公开了一种电沉积壳聚糖-离子液体-酶复合膜制备修饰电极的方法，其特征在于：制备修饰时将电极作为阴极置入含壳聚糖、离子液体和酶的酸性溶液中，通电进行电沉积，壳聚糖、离子液体和酶附着在阴电极表面，得到修饰电极。本发明电沉积壳聚糖-离子液体-酶复合膜制备修饰电极的方法，条件温和，简便快捷，壳聚糖-离子液体-酶复合膜厚度可控，并可根据需要控制酶固定量。The invention discloses a method for preparing a modified electrode by electrodepositing a chitosan-ionic liquid-enzyme composite membrane, which is characterized in that the electrode is used as a cathode and placed in an acidic solution containing chitosan, ionic liquid and enzyme when preparing the modification , electrified for electrodeposition, chitosan, ionic liquid and enzyme attached to the surface of the negative electrode to obtain a modified electrode. The method for preparing a modified electrode by electrodepositing a chitosan-ionic liquid-enzyme composite film of the invention has mild conditions, is simple and quick, and the thickness of the chitosan-ionic liquid-enzyme composite film is controllable, and the amount of enzyme immobilization can be controlled as required.
Description
Technical field
The present invention relates to the modified electrode preparation, relate in particular to the method that a kind of electro-deposition chitosan-ionic liquid-enzyme compound film prepares modified electrode.
Background technology
Biology sensor is the novel sensor technology that grows up nearly decades, has advantages such as analysis speed is fast, easy to use, has significant application value in fields such as biomedicine, clinical diagnosis, Industry Control, environmental monitoring, food medical industries.Enzyme modified electrode is the focus of biology sensor research, and has developed multiple electrode preparation method.The sensitivity of enzyme modified electrode and stability are to estimate two important indicators of enzyme electrode performance, and the controllability of enzyme modified electrode preparation process is the principal element of decision enzyme electrode practical application.The selection of enzyme fixing means and electron transfer promoter is fast, highly sensitive for preparation response, the biology sensor of good stability, long service life is most important.
Ionic liquid is the novel green solvent of a class.Compare with organic solvent, ionic liquid has many good performances, as high ionic conductivity; The electrochemical window of broad; Steam forces down, and is not volatile etc.Along with deepening continuously of ionic liquid application in recent years, it is found that many enzymes have good performance in ionic liquid, help carrying out galvanochemistry and electrocatalytic reaction.Simultaneously, the ionic conductivity of ionic liquid height also can effectively promote the electron transport between enzyme and electrode material.
Modified electrode is as the vitals in the biology sensor, and its preparation method has a significant impact the performance of biology sensor.In the prior art about modified electrode preparation as:
Chinese invention patent application 200610017201.8 discloses the method that a kind of ion liquid sol-gel compound membrane buried enzyme prepares biology sensor, its key step is: with the 1-butyl, 3-methylimidazole tetrafluoride boron, tetraethoxy-silicane, water, the hydrochloric acid of 0.1mol/L at room temperature mixes according to the volume ratio of 1-2: 2-4: 1-2: 0.05-0.1, magnetic agitation 2-5 hour, placed 1-2 hour under the room temperature, obtain Sol A; Is 1-3 with the phosphate buffer solution of the pH=6-7 of enzyme and 0.05mol/L according to mass volume ratio: the ratio of 50 milligrams of microlitres mix solution B; With Sol A and solution B according to volume ratio 1: 1-2.5 mixes, and gets this mixed liquor and evenly is coated to electrode surface, and thickness is the 0.5-500 micron, placed 24-48 hour down in 0-4 ℃, biology sensor.
But enzyme modified electrode preparation technology is generally complicated in the prior art, and manufacturing cycle is long.
Summary of the invention
The present invention provides a kind of mild condition, and the controlled electro-deposition chitosan-ionic liquid-enzyme compound film of film thickness prepares the method for modified electrode, mainly is the improvement at modified electrode preparation method in the biology sensor.
A kind of electro-deposition chitosan-ionic liquid-enzyme compound film prepares the method for modified electrode, insert electrode in the acid solution of chitosan-containing, ionic liquid and enzyme as negative electrode during the preparation modified electrode, electro-deposition is carried out in energising, shitosan, ionic liquid and enzyme obtain modified electrode attached to the negative electrode surface.
It is as follows that method is put in the preparation of described acid solution:
A) shitosan being dissolved in acetate or the mass percent concentration that mass percent concentration is 0.5-5.0% is in the hydrochloric acid solution of 0.5-5.0%, obtains solution A;
The mass percent concentration of shitosan is 0.5-4.0% in the solution A;
B) in solution A, add ionic liquid, after magnetic agitation 1-3 hour, ultrasonic 10-30 minute, obtain solution B;
Described ionic liquid is 1-butyl 3-methylimidazole tetrafluoride boron or 1-butyl 3-methylimidazole chlorine, and the ionic liquid quality is the 1-3% of solution A quality;
C) in solution B, add enzyme, ultrasonic 10-30 minute, obtain acid solution;
Described enzyme is one or more in peroxidase, glucose oxidase, Lactate Oxidase, cholesterol oxidase, Tyrosine oxidase, alcohol oxidase, alcohol dehydrogenase, laccase, CHE, choline oxidase, bilirubin oxidase, lipase or the polyphenol oxidase, and the concentration of enzyme in acid solution is 0.01-10mg/mL.
Described electrode is gold, platinum, glass carbon, tin indium oxide.
Described electrodeposition time is 1-20 minute, the negative electrode current potential is 0～-2V.
The shitosan big molecular biosciences polysaccharide of straight chain that to be natural class polysaccharide chitin obtain after partially deacetylated, chemistry poly-(1,4)-2-amino by name-2-deoxidation-callose.The shitosan source is abundant, is the environment renewable resource, has excellent biological compatibility.Active aminos a large amount of in the chitosan molecule can be protonated under sour environment, makes shitosan be dissolved in acid solution.
The present invention is compound as a kind of novel enzyme fixation support with shitosan and ionic liquid.Insert electrode in the mixed solution of shitosan, ionic liquid and enzyme, during energized, the hydrogen ion in the solution is reduced into hydrogen at negative electrode.Meanwhile, cathode surface pH value increases gradually, and forms certain pH gradient in solution.Because the dissolubility of shitosan is relevant with solution acidity, when the pH value of solution value was higher than shitosan pKa near the electrode, shitosan became insoluble state and is deposited on cathode surface.Shitosan is combined with electron transfer promoter ionic liquid and enzyme, promptly in chitosan solution, add ionic liquid and enzyme, can obtain shitosan-ionic liquid-enzyme composite membrane in the electrode surface electro-deposition, the preparation enzyme modified electrode.
Prepare methods such as modified electrode compares with conventional drop-coating, electro-deposition chitosan-ionic liquid-enzyme compound film of the present invention prepares the method for modified electrode, and mild condition is simple and efficient, shitosan-ionic liquid-enzyme composite membrane controllable thickness, and can control the enzyme fixed amount as required.
Shitosan has excellent biological compatibility, is good enzyme immobilization carrier.Many enzymes also have good performance in ionic liquid.Simultaneously, the ionic conductivity of ionic liquid height can effectively promote the electron transport between enzyme and electrode material.Shitosan-ionic liquid-enzyme composite membrane can make enzyme have good biologically active and be easy to take place galvanochemistry and electrocatalytic reaction.The modified electrode response of the shitosan-ionic liquid of the inventive method preparation-enzyme composite membrane preparation is fast, highly sensitive, good stability, long service life.
Embodiment
Embodiment 1
It is in 1% the acetic acid solution that shitosan is dissolved in mass percent concentration, the mass percent concentration of shitosan is 0.5%, add ionic liquid 1-butyl 3-methylimidazole tetrafluoride boron, the ionic liquid quality is 1% of a chitosan solution quality, magnetic agitation 3 hours, ultrasonic dispersion adds 1mg/mL horseradish peroxidase, ultrasonic dispersion 15 minutes after 30 minutes.As negative electrode, electro-deposition is 5 minutes under-1V current potential, promptly gets the complex film modified electrode of electro-deposition shitosan-ionic liquid-horseradish peroxidase with gold electrode, and the modified electrode that obtains can be used in the biology sensor.
Embodiment 2
It is in 1% the hydrochloric acid solution that shitosan is dissolved in mass percent concentration, the mass percent concentration of shitosan is 1%, add ionic liquid 1-butyl 3-methylimidazole tetrafluoride boron, the ionic liquid quality is 2% of a chitosan solution quality, magnetic agitation 1 hour, ultrasonic dispersion adds 2mg/mL glucose oxidase, ultrasonic dispersion 10 minutes after 10 minutes.As negative electrode, electro-deposition is 8 minutes under-1.2V current potential, promptly gets the complex film modified electrode of electro-deposition shitosan-ionic liquid-glucose oxidase with glass-carbon electrode, and the modified electrode that obtains can be used in the biology sensor.
Embodiment 3
It is in 0.5% the acetic acid solution that shitosan is dissolved in mass percent concentration, the mass percent concentration of shitosan is 1%, add ionic liquid 1-butyl 3-methylimidazole chlorine, the ionic liquid quality is 1% of a chitosan solution quality, magnetic agitation 3 hours, ultrasonic dispersion adds 1mg/mL glucose oxidase and 2mg/mL horseradish peroxidase, ultrasonic dispersion 10 minutes after 30 minutes.As negative electrode, electro-deposition is 5 minutes under-1.5V current potential, promptly gets electro-deposition shitosan-ionic liquid-two enzyme composite membrane modified electrodes with platinum electrode, and the modified electrode that obtains can be used in the biology sensor.
Claims (5)
1. an electro-deposition chitosan-ionic liquid-enzyme compound film prepares the method for modified electrode, it is characterized in that: insert electrode in the acid solution of chitosan-containing, ionic liquid and enzyme as negative electrode, electro-deposition is carried out in energising, shitosan, ionic liquid and enzyme obtain modified electrode attached to the negative electrode surface;
Described ionic liquid is 1-butyl 3-methylimidazole tetrafluoride boron or 1-butyl 3-methylimidazole chlorine;
Described acid solution is that mass percent concentration is that acetate or the mass percent concentration of 0.5-5.0% is the hydrochloric acid solution of 0.5-5.0%.
2. the method for claim 1, it is characterized in that: the compound method of described acid solution is as follows:
A) shitosan is dissolved in the hydrochloric acid solution of acetate that mass percent concentration is 0.5-5.0% or 0.5-5.0%, obtains solution A; The mass percent concentration of shitosan is 0.5-4.0% in the solution A;
B) in solution A, add ionic liquid, after magnetic agitation 1-3 hour, ultrasonic 10-30 minute, obtain solution B;
Described ionic liquid quality is the 1-3% of solution A quality;
C) in solution B, add enzyme, ultrasonic 10-30 minute, obtain acid solution;
Described enzyme is one or more in peroxidase, glucose oxidase, Lactate Oxidase, cholesterol oxidase, Tyrosine oxidase, alcohol oxidase, alcohol dehydrogenase, laccase, CHE, choline oxidase, bilirubin oxidase, lipase or the polyphenol oxidase, and the concentration of enzyme in acid solution is 0.01-10mg/mL.
3. the method for claim 1 is characterized in that: described electrode is gold, platinum, glass carbon, tin indium oxide.
4. the method for claim 1, it is characterized in that: described electrodeposition time is 1-20 minute.
5. the method for claim 1 is characterized in that: during described electro-deposition the negative electrode current potential be 0～-2V.
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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101308112B (en) * | 2008-07-07 | 2011-07-20 | 浙江大学 | Electrodeposit method for preparing modification electrode of chitosan -dye-enzyme composite film |
| CN101931082B (en) * | 2009-06-24 | 2013-02-20 | 中国科学院化学研究所 | High interference resistance enzyme biofuel battery cathode and preparation method thereof |
| CN103013952B (en) * | 2012-01-16 | 2014-07-23 | 南京工业大学 | Functional ionic liquid modified lipase and modification method thereof |
| CN102559635B (en) * | 2012-01-16 | 2013-04-10 | 南京工业大学 | Functional ionic liquid modified lipase and modification method thereof |
| CN102636540A (en) * | 2012-04-19 | 2012-08-15 | 湖南大学 | Glucose detection sensor, and preparation and application methods thereof |
| CN103543187A (en) * | 2012-07-12 | 2014-01-29 | 西北师范大学 | Hydrophobic ionic liquid fixed cholesterol enzyme/Prussian blue modified electrode for detecting cholesterol |
| CN105755523B (en) * | 2016-05-09 | 2017-11-21 | 扬州大学 | A kind of preparation method of hydrophobe plasma membrane |
| CN106383160A (en) * | 2016-12-09 | 2017-02-08 | 北京化工大学 | Ionic liquid modified urate oxidase enzyme membrane and preparation method thereof |
| CN115290735A (en) * | 2022-08-19 | 2022-11-04 | 中国科学院深圳先进技术研究院 | An enzyme-modified electrochemical electrode, a preparation method thereof, and an ultra-thin enzyme plating device |
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| CN1309195A (en) * | 2000-12-07 | 2001-08-22 | 厦门大学 | Bioactive composite Ca-P ceramics/chitosan film on metal surface and electrochemical co-deposition process for preparing it |
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