CN102944597B - Deliquescent-polyelectrolyte-based full-solid-state ethanol gas sensor enzyme electrode and manufacturing method thereof - Google Patents
Deliquescent-polyelectrolyte-based full-solid-state ethanol gas sensor enzyme electrode and manufacturing method thereof Download PDFInfo
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Abstract
The invention relates to a deliquescent-polyelectrolyte-based full-solid-state ethanol gas sensor enzyme electrode and a manufacturing method thereof. The manufacturing method comprises the following steps: on a matrix made of an insulating material, preparing a printed electrode composed of a working electrode, a reference electrode and a counter electrode by screen printing technology; coating a polycarbonate insulator on the surfaces in the middle; coating a reaction layer composed of graphene, deliquescent polyelectrolyte and alcohol oxidase on the working electrode; and coating a deliquescent solid polymer film among the working electrode, counter electrode and reference electrode to obtain the biological enzyme electrode. The invention provides an enzyme electrode for gas-state ethanol detection; and the enzyme electrode can implement high-selectivity detection of ethanol gas without using any liquid electrolyte in the use process, thereby integrally enhancing the properties and practicality of the ethanol gas biosensor.
Description
Technical field
The present invention relates to a kind of all solid state alcohol gas sensor enzyme electrode based on deliquescence type polyelectrolyte and preparation method thereof.
Background technology
Along with the mankind are to the attention degree of self health with the concern of ecologic environment is deepened day by day, Monitoring and Controlling to gas is had higher requirement, this research that is gas sensor, development and production provide opportunities and challenges, also to gas sensor, have proposed new research topic.In order to meet these requirements, gas sensor must have higher sensitivity and selectivity, and repeatability and good stability, can manufacture, cost performance high.But traditional gas sensor is due to the restriction of itself material, the specific selectivity of pure gas sensor is all poor, only has seldom several gas sensor arrays, using the method for neural network as late time data disposal system, can realize to a certain extent the selectivity identification to gas, even so its overall performance still depends on the performance of single-sensor element, and the selectivity of sensor array is greatly limited.Therefore, improve gas sensor specificity, significant for applying of gas sensor.
The most outstanding feature of biology sensor is specificity, and biological identification molecule is combined with gas sensor, will be expected to obtain the gas sensor of high specific.In biogas sensor research, the enzymic catalytic reaction also mainly depending on gas/liquid interface at present carries out (Anal.Chem, (1994), 66,3297-3302; Sens.Actuators B, (2002), 83, (1-3), 35-40; Sens.Actuators B, (2005), 108, (1-2), 639-645; Sens.Actuators B, (2000), 70, (1-3), 182-187; Biosens. & Bioelectron., (2002), 17, (5), 389-394; Biosens.Bioelectron., (2002), 17, (5), 427-432; Sens.Actuators B, (2001), 81, (1), 107-114; Biosens.Bioelectron., (2002), 17, (9), 789-796), really not realizing the detection of gaseous sample in gas phase, inevitably can there is electrolyte evaporation or leakage in these methods, decay (the Sens.Actuators B that causes sensor signal, (2005), 107,812-817), and device preparation is complicated, be unfavorable for microminiaturization, intelligent and integrated.We utilize serigraphy three-electrode system in conjunction with solid polymer electrolyte, built all solid state alcohol gas sensor (20111046159.8), realized the detection of alcohol gas on gas/solid interface, but the response of biogas sensor is still subject to the restriction of moisture, utilization of the present invention has fixation support and the solid electrolyte that the polyelectrolyte of deliquescence character is enzyme, maintain the catalytic activity that enzyme molecule is higher, charge transfer in the process of realization response simultaneously, has built new bio gas sensor.
Summary of the invention
The deficiency existing in detection in order to overcome existing alcohol gas biology sensor, the object of this invention is to provide a kind of all solid state alcohol gas sensor enzyme electrode based on deliquescence type polyelectrolyte and preparation method thereof, the enzyme electrode making by the method has improved properties and the practicality of biology sensor on the whole.
The technical solution adopted in the present invention is as follows:
The enzyme electrode that all-solid ethanol gas biosensor of the present invention is used has insulating material matrix, at matrix surface screen printing, be brushed with working electrode, contrast electrode and to electrode, in the middle of electrode, apply one deck carbonic ester insulator, on working electrode, be coated with the responding layer being formed by Graphene, deliquescence type polyelectrolyte and alcohol oxidase, then at working electrode, contrast electrode with between to electrode, apply deliquescence type polyelectrolyte membrane.
The method for making of the enzyme electrode that all-solid ethanol gas biosensor is used, comprises the following steps:
1) preparation of deliquescence type polyelectrolyte
A. in 100mL there-necked flask, according to 1: 1 (mol: ratio mol) joins diethylenetriamine and methyl acrylate in a certain amount of methyl alcohol, reaction 12h; Then under decompression state, be warming up to 30 ℃ of reaction 1h, at 60 ℃, react 1h, at 100 ℃, react 2h, at 120 ℃, react 3h, Depressor response 2h at 140 ℃, after reaction finishes, dilutes with absolute ethyl alcohol, and wash, precipitate in a large amount of ether, vacuum drying, obtains ultrabranching polyamide;
B. in 100mL there-necked flask, by ultrabranching polyamide with deionized water, formic acid, formaldehyde according to 1: 10: 7: the ratio of 7 (g/g) is mixed, and 75 ℃ of back flow reaction certain hours, then repeatedly wash, precipitate with ether, vacuum drying;
C. the product in above-mentioned b is dissolved in 50mL methyl alcohol, then adds a certain amount of halogenated alkane, 50 ℃ of reaction certain hours, product dissolves through methyl alcohol, and ether sedimentation is repeatedly purified, and vacuum drying obtains deliquescence type polyelectrolyte.
2) clean surface has working electrode, contrast electrode and to the pottery of electrode or PVC base system, dry for standby;
3) glutaraldehyde that is 2% by the Graphene of the alcohol oxidase of 3～6mg/mL, 0.5～3mg/mL, 5～20mg/mL deliquescence type polyelectrolyte and volume ratio concussion mixes, then getting 3～10 μ L drips and is coated in working electrode surface, naturally dry, form responding layer;
4) the poly-electrolytic solution of the deliquescence type that is 2～5% by volume ratio is evenly coated in working electrode, contrast electrode and between electrode, after naturally drying, obtains enzyme electrode.
Advantage of the present invention is:
1) choose working electrode, contrast electrode and electrode is printed on to the three-electrode system on same matrix, being convenient to microminiaturization, the intellectuality and integrated of sensor electrode;
2) use of Graphene has improved the response sensitivity of sensor;
3) alcohol oxidase, as the recognition component of ethanol molecule, has improved the response selectivity of sensor greatly;
4) utilize solid-state polyelectrolyte as the electric charge transport vehicle in electrochemical reaction process, avoid using liquid electrolyte, improve the stability of sensor signal;
5) deliquescence type polyelectrolyte, as the fixation support of enzyme, can be enzymic catalytic reaction sufficiently high water activity is provided, and significantly improves response signal;
6) to have volume little for enzyme electrode of the present invention, low cost, the advantage such as easy to use.And response sensitivity is high, stability is better, and response selectivity is high, can be used for accurate measurement and the control of ethanol gas concentration.
Accompanying drawing explanation
Fig. 1 is the structural representation of enzyme electrode of the present invention.
Fig. 2 is the deliquescence procedure chart of the deliquescence type polyelectrolyte of the embodiment of the present invention 1 making, A: drying regime, B:40% relative humidity deliquescence 1h, C:40% relative humidity deliquescence 2h, D: state of saturation.
Fig. 3 is the deliquescence procedure chart that the embodiment of the present invention 2 is made deliquescence type polyelectrolyte, A: drying regime, B:40% relative humidity deliquescence 1h, C:40% relative humidity deliquescence 2h, D: state of saturation.
Fig. 4 is the response curve of the enzyme electrode made of the embodiment of the present invention 2 to alcohol vapour.
Embodiment
Below in conjunction with drawings and Examples, further illustrate the present invention:
With reference to accompanying drawing 1, the enzyme electrode that all-solid ethanol gas biosensor is used has insulating body (8), at matrix surface screen printing, be brushed with working electrode (1), contrast electrode (2) and to electrode (3), on their center section face, scribble one deck coated electrode table and polycarbonate insulator (4).Working electrode (1), contrast electrode (2) and electrode (3) is corresponded respectively to pin (5), (6) and (7) of base end.Working electrode is coated with the responding layer being comprised of bioactive materials, nano material, alcohol oxidase on (1), then at working electrode (1), contrast electrode (2) with between to electrode (3), applies polyelectrolyte membrane.
Described working electrode (1) is circular, and diameter is 2～4mm, and material is platinum, and contrast electrode is silver/silver chloride, to electrode, is platinum.
Embodiment 1:
1) preparation of deliquescence type polyelectrolyte
A. in 100mL there-necked flask, the methyl acrylate of the diethylenetriamine of 0.2mol and 0.2mol is joined in 25mL methyl alcohol to reaction 12h; Then under decompression state, be warming up to 30 ℃ of reaction 1h, at 60 ℃, react 1h, at 100 ℃, react 2h, at 120 ℃, react 3h, Depressor response 2h at 140 ℃, after reaction finishes, dilutes with absolute ethyl alcohol, and wash, precipitate in a large amount of ether, vacuum drying, obtains ultrabranching polyamide;
B. in 100mL there-necked flask, 5g ultrabranching polyamide is mixed with 50mL deionized water, 35g formic acid and 35g formaldehyde, 75 ℃ of back flow reaction certain hours, then repeatedly wash, precipitate vacuum drying with ether;
C. the product in above-mentioned b is dissolved in 50mL methyl alcohol, then adds 10.32g bromination of n-butane, 50 ℃ of reaction 48h, product dissolves through methyl alcohol, and ether sedimentation is repeatedly purified, and vacuum drying obtains deliquescence type polyelectrolyte.
2) clean surface has working electrode (diameter 4mm), silver/silver chloride contrast electrode and the ceramic matrix of platinum to electrode, dry for standby;
3) glutaraldehyde that is 2% by the Graphene of the alcohol oxidase of 6mg/mL, 1mg/mL, 10mg/mL deliquescence type polyelectrolyte and volume ratio concussion mixes, and then gets 6 μ L and drips and be coated in working electrode surface, naturally dries formation responding layer;
4) the poly-electrolytic solution of the deliquescence type that is 3% by volume ratio is evenly coated in working electrode, contrast electrode and between electrode, after naturally drying, obtains enzyme electrode.
Embodiment 2:
1) preparation of deliquescence type polyelectrolyte
A. in there-necked flask, the methyl acrylate of the diethylenetriamine of 0.4mol and 0.4mol is joined in 50mL methyl alcohol to reaction 12h; Then under decompression state, be warming up to 30 ℃ of reaction 1h, at 60 ℃, react 1h, at 100 ℃, react 2h, at 120 ℃, react 3h, Depressor response 2h at 140 ℃, after reaction finishes, dilutes with absolute ethyl alcohol, and wash, precipitate in a large amount of ether, vacuum drying, obtains ultrabranching polyamide;
B. in there-necked flask, 10g ultrabranching polyamide is mixed with 100mL deionized water, 70g formic acid and 70g formaldehyde, 75 ℃ of back flow reaction certain hours, then repeatedly wash, precipitate with ether, vacuum drying;
C. the product in above-mentioned b is dissolved in 100mL methyl alcohol, then adds 24.86g bromo normal hexane, 50 ℃ of reaction 24h, product dissolves through methyl alcohol, and ether sedimentation is repeatedly purified, and vacuum drying obtains deliquescence type polyelectrolyte.
2) clean surface has working electrode (diameter 4mm), silver/silver chloride contrast electrode and the ceramic matrix of platinum to electrode, dry for standby;
3) glutaraldehyde that is 2% by the Graphene of the alcohol oxidase of 3mg/mL, 0.5mg/mL, 5mg/mL deliquescence type polyelectrolyte and volume ratio concussion mixes, and then gets 10 μ L and drips and be coated in working electrode surface, naturally dries formation responding layer;
4) the poly-electrolytic solution of the deliquescence type that is 2% by volume ratio is evenly coated in working electrode, contrast electrode and between electrode, after naturally drying, obtains enzyme electrode.
Embodiment 3:
1) preparation of deliquescence type polyelectrolyte
A. in there-necked flask, the methyl acrylate of the diethylenetriamine of 0.2mol and 0.2mol is joined in 25mL methyl alcohol, under agitation slowly drip the acrylic acid first vinegar of 0.2mol, reaction 12h; Then under decompression state, be warming up to 30 ℃ of reaction 1h, at 60 ℃, react 1h, at 100 ℃, react 2h, at 120 ℃, react 3h, Depressor response 2h at 140 ℃, after reaction finishes, dilutes with absolute ethyl alcohol, and wash, precipitate in a large amount of ether, vacuum drying, obtains ultrabranching polyamide;
B. in there-necked flask, 5g ultrabranching polyamide is mixed with 50mL deionized water, 35g formic acid and 35g formaldehyde, 75 ℃ of back flow reaction certain hours, then repeatedly wash, precipitate with ether, vacuum drying;
C. the product in above-mentioned b is dissolved in 50mL methyl alcohol, then adds 10.32g bromination of n-butane, 50 ℃ of reaction 72h, product dissolves through methyl alcohol, and ether sedimentation is repeatedly purified, and vacuum drying obtains deliquescence type polyelectrolyte.
2) clean surface has working electrode (diameter 2mm), silver/silver chloride contrast electrode and the ceramic matrix of platinum to electrode, dry for standby;
3) glutaraldehyde that is 2% by the Graphene of the alcohol oxidase of 5mg/mL, 1mg/mL, 5mg/mL deliquescence type polyelectrolyte and volume ratio concussion mixes, and then gets 8 μ L and drips and be coated in working electrode surface, naturally dries formation responding layer;
4) the poly-electrolytic solution of the deliquescence type that is 3% by volume ratio is evenly coated in working electrode, contrast electrode and between electrode, after naturally drying, obtains enzyme electrode.
Claims (4)
1. the method for making of all solid state alcohol gas sensor enzyme electrode based on deliquescence type polyelectrolyte, described electrode has insulating material matrix, at matrix surface screen printing, be brushed with working electrode, contrast electrode and to electrode, in the middle of electrode, apply one deck carbonic ester insulator, on working electrode, be coated with by Graphene, the responding layer that deliquescence type polyelectrolyte and alcohol oxidase form, then at working electrode, contrast electrode and to applying deliquescence type polyelectrolyte membrane between electrode, described insulating material matrix is Polyvinylchloride or pottery, electrode adopts screen printing technique to print, described working electrode is circular, diameter is 2～4mm, material is platinum, contrast electrode is silver/silver chloride, to electrode, be platinum, it is characterized in that: described method for making comprises the following steps:
1) preparation of deliquescence type polyelectrolyte
A. in there-necked flask, according to the ratio of 1: 1, diethylenetriamine and methyl acrylate are joined in a certain amount of methyl alcohol, stir lower reaction 12h; Then under decompression state, be warming up to 30 ℃ of reaction 1h, at 60 ℃, react 1h, at 100 ℃, react 2h, at 120 ℃, react 3h, Depressor response 2h at 140 ℃, after reaction finishes, dilutes with absolute ethyl alcohol, and wash, precipitate in a large amount of ether, vacuum drying, obtains ultrabranching polyamide;
B. in there-necked flask, by ultrabranching polyamide with deionized water, formic acid, formaldehyde according to 1: 10: 7: the ratio of 7 (g/g) is mixed, and 75 ℃ of back flow reaction certain hours, then repeatedly wash, precipitate with ether, vacuum drying;
C. the product in above-mentioned b is dissolved in 50mL methyl alcohol, then adds a certain amount of halogenated alkane, 50 ℃ of reaction certain hours, product dissolves through methyl alcohol, and ether sedimentation is repeatedly purified, and vacuum drying obtains deliquescence type polyelectrolyte,
2) clean surface has working electrode, contrast electrode and to the pottery of electrode or PVC base system, dry for standby;
3) the glutaraldehyde concussion that is 2% by the deliquescence type polyelectrolyte of the Graphene of the alcohol oxidase of 3～6mg/mL, 0.5～3mg/mL, 5～20mg/mL and volume ratio mixes, then getting 3～10 μ L drips and is coated in working electrode surface, naturally dry, form responding layer;
4) the poly-electrolytic solution of the deliquescence type that is 2～5% by volume ratio is evenly coated in working electrode, contrast electrode and between electrode, after naturally drying, obtains enzyme electrode.
2. the method for making of a kind of all solid state alcohol gas sensor enzyme electrode based on Graphene according to claim 1, is characterized in that: halogenated alkane used is a kind of in bromination of n-butane, bromo normal hexane.
3. the method for making of a kind of all solid state alcohol gas sensor enzyme electrode based on Graphene according to claim 1, is characterized in that: in the preparation of deliquescence type polyelectrolyte, the reaction time of b step is 6～12h.
4. the method for making of a kind of all solid state alcohol gas sensor enzyme electrode based on Graphene according to claim 1, is characterized in that: in the preparation of deliquescence type polyelectrolyte, the reaction time of c step is 6～72h.
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| US20100092809A1 (en) * | 2008-10-10 | 2010-04-15 | Board Of Trustees Of Michigan State University | Electrically conductive, optically transparent films of exfoliated graphite nanoparticles and methods of making the same |
| CN101818139A (en) * | 2010-04-22 | 2010-09-01 | 无锡市金坤生物工程有限公司 | Preparation method of immobilization peroxidase |
| CN102323300A (en) * | 2011-07-18 | 2012-01-18 | 浙江大学 | Polyelectrolyte and graphene composite resistive moisture sensor and manufacturing method thereof |
| CN102495118A (en) * | 2011-12-14 | 2012-06-13 | 天津工业大学 | Enzyme electrode for all-solid ethanol gas biosensor and method for manufacturing enzyme electrode |
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| US20100092809A1 (en) * | 2008-10-10 | 2010-04-15 | Board Of Trustees Of Michigan State University | Electrically conductive, optically transparent films of exfoliated graphite nanoparticles and methods of making the same |
| CN101818139A (en) * | 2010-04-22 | 2010-09-01 | 无锡市金坤生物工程有限公司 | Preparation method of immobilization peroxidase |
| CN102323300A (en) * | 2011-07-18 | 2012-01-18 | 浙江大学 | Polyelectrolyte and graphene composite resistive moisture sensor and manufacturing method thereof |
| CN102495118A (en) * | 2011-12-14 | 2012-06-13 | 天津工业大学 | Enzyme electrode for all-solid ethanol gas biosensor and method for manufacturing enzyme electrode |
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| 苗智颖，李莎，秦霞，王会才，陈强.多壁碳纳米管增效层层自组装构建乙醇传感器.《传感器与微***》.2010,第29卷(第12期),85-87. * |
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