CN109030602B - A kind of electric potential type organophosphorus biosensor and preparation method thereof - Google Patents
A kind of electric potential type organophosphorus biosensor and preparation method thereof Download PDFInfo
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- CN109030602B CN109030602B CN201810906777.2A CN201810906777A CN109030602B CN 109030602 B CN109030602 B CN 109030602B CN 201810906777 A CN201810906777 A CN 201810906777A CN 109030602 B CN109030602 B CN 109030602B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
Abstract
The invention discloses a kind of electric potential type organophosphorus biosensors and preparation method thereof.The biosensor is used for the quick detection of organophosphorus pesticide.Acetylcholinesterase is fixed on the organic nucleopore membranes basis material of polyethylene terephthalate by the present invention using Deacetyled konjac glucomannan colloidal sol investment; the enzyme membrane of preparation is fixed in the glass electrode of pH meter with ' O ' shape circle again, obtains organophosphorus pesticide biosensor.The organophosphorus pesticide sensor of this method preparation has the characteristics that high sensitivity, response are fast, at low cost.
Description
Technical field
The present invention relates to a kind of electric potential type organophosphorus biosensors and preparation method thereof, belong to analysis and testing technology neck
Domain.
Background technique
The event that pesticide residue causes people, animal to be poisoned happens occasionally, and the hidden danger such as carcinogenic, teratogenesis and cause gene mutation more enable
People's worry.Organophosphorus pesticide is that use scope is most wide, a kind of maximum pesticide of usage amount.Current organophosphorus pesticide detection side
Method be mainly chromatography [Wang Zongxian, Gao Zhixian, Ma Cheng, the progress of organic phosphorus detection method, China Health, 2003,13 (4),
401-403].It is high to the technical requirements of testing staff since chromatography pretreatment process is cumbersome, be not suitable for field quick detection.
Biosensor technology based on electrochemical analysis principle has quick, sensitive, accurate and is easy to continuous and automatic
The characteristics of, it can satisfy the needs of field quick detection.In existing electric potential type organophosphorus pesticide biosensor, it is common that
Acetylcholinesterase is fixed on nylon wire using cross-linking method or covalent bonding method, is then fixed nylon wire with ' O ' shape circle
On to the glass electrode of pH meter [immobilization research of the acetylcholinesterase on nylon wire, chemistry world, 2007,48 (12):
751-753；The research of acetylcholinesterasebiosensor biosensor, pesticide, 2008,47 (8): 577-579].The enzyme membrane being prepared
Unevenly, and the compactness of enzyme membrane and electrode is not high, thus the sensor consistency and repeatability prepared are not ideal enough.
Summary of the invention
For overcome the deficiencies in the prior art, the purpose of the present invention is to provide a kind of electric potential type organophosphorus biosensors
And preparation method thereof.Acetylcholinesterase is fixed in the organic nucleopore membranes of polyethylene terephthalate by the present invention, preparation
Sensor structure out is stablized, consistency and reproducible.
Acetylcholinesterase is fixed on poly- to benzene two by the present invention using Deacetyled konjac glucomannan colloidal sol investment
On the organic nucleopore membranes basis material of formic acid glycol ester, then the enzyme membrane of preparation is fixed on to the glass electrode of pH meter with ' O ' shape circle
On, obtain electric potential type organophosphorus biosensor.Technical solution of the present invention is specifically described as follows.
The present invention provides a kind of preparation method of electric potential type organophosphorus biosensor, comprising the following steps:
(1) preparation of Deacetyled konjac glucomannan colloidal sol
It is 0.8~1.2g/L's by 0.8~1.2g Deacetyled konjac glucomannan fine powder and 80~120ml concentration
Na2CO3After solution is mixed into translucent colloidal liquid, it is further continued for 15~25min of stirring, it is sweet that deacetylated konjac Portugal is made
Glycan colloidal sol；
(2) preparation of enzyme membrane
The acetylcholinesterase, 0.8~1.2ml phosphate buffer solution and 0.8~1.2ml for weighing 0.8~1.2mg are deacetylated
Base KGM Sol is uniformly mixed, obtains mixed solution；Take 2~5 μ L mixing drop-coated in poly terephthalic acid second two
In the organic nucleopore membranes of alcohol ester, then it is placed in refrigerator freeze-drying lyophilization at a temperature of 4 DEG C, obtains enzyme membrane；
(3) preparation of enzyme electrode
Enzyme membrane is fixed on the combination electrode of pH meter with ' O ' shape circle, enzyme electrode is made, is i.e. the organic phosphorus biology of electric potential type passes
Sensor.
In the present invention, in step (2), the aperture of the organic nucleopore membranes of polyethylene terephthalate is 1-10 μm.
The present invention also provides electric potential type organophosphorus biosensors made from a kind of above-mentioned preparation method.
Compared to the prior art, the beneficial effects of the present invention are:
Acetylcholinesterase is fixed on poly terephthalic acid using Deacetyled konjac glucomannan investment by the present invention
In the organic nucleopore membranes of glycol ester, enzyme membrane is obtained with good hydrophilic property, good penetrability, flat and smooth, high mechanical strength, flexibility
The advantages that good.And then the Copper diethlydithiocarbamate prepared using the enzyme membrane, the transducer sensitivity is high, response is fast, consistency and
Reproducible advantage.
Detailed description of the invention
Fig. 1 is pesticide to the inhibiting rate of the sensor potential response value in embodiment 1 and the logarithm relationship of pesticide concentration.
Specific embodiment
The present invention is described in further detail below by embodiment, but is not intended to limit the present invention.
Embodiment 1
1. the preparation of Deacetyled konjac glucomannan colloidal sol
It weighs 0.8g Deacetyled konjac glucomannan fine powder and is added to the Na that 100ml concentration is 1.0g/L2CO3In solution,
It is stirred simultaneously with electric blender.Become translucent colloidal liquid in solution, stirs 20min with electric blender again at this time.It will
The colloidal sol of preparation is transferred in bottle,suction, is filtered bottleneck and is blocked with rubber stopper, is evacuated to bubble-free in colloidal sol.
2. the preparation of enzyme membrane
The acetylcholinesterase of 1mg is weighed in centrifuge tube, and 1ml phosphate buffer solution is added, it is deacetylated to add 1ml
Base KGM Sol is uniformly mixed.Take the poly terephthalic acid second two that the above-mentioned mixing drop-coated of 2 μ l is 10 μm in aperture
In the organic nucleopore membranes of alcohol ester, it is placed in 4 DEG C of freeze-drying lyophilizations in refrigerator.
3. the preparation of enzyme electrode
The enzyme membrane prepared is fixed on the combination electrode of pH meter with ' O ' shape circle, obtains enzyme electrode.
Embodiment 2
With embodiment 1, difference is only that the organic nucleopore membranes of polyethylene terephthalate for other implementation conditions and parameter
Aperture be 5 μm.
Embodiment 3
With embodiment 1, difference is only that it is the organic nucleopore of polyethylene terephthalate for other implementation conditions and parameter
The aperture of film is 1 μm.
Embodiment 4
With embodiment 1, difference is only that the Deacetyled konjac glucomannan colloidal sol of drop coating for other implementation conditions and parameter
Volume with the mixed liquor of acetylcholinesterase is 3 μ l.
Embodiment 5
With embodiment 1, difference is only that the Deacetyled konjac glucomannan colloidal sol of drop coating for other implementation conditions and parameter
Volume with the mixed liquor of acetylcholinesterase is 5 μ l.
Application Example 1
By organophosphorus pesticide biosensor assay organophosphorus pesticide DDVP made from embodiment 1.Temperature is 30 DEG C, bottom
Object (acecoline solution) concentration is 8 × 10-3mol·L-1, in pH=7.0 buffer solution, incubation of the enzyme membrane in pesticide
Time is 3min.
Pesticide is as shown in Figure 1 to the inhibiting rate of sensor potential response value and the logarithm relationship of pesticide concentration.As a result table
Bright, linear response range is 1~10mg/L, and Monitoring lower-cut is up to 0.4mg/L.
The test-strips for extracting 10 batches test same actual sample, test relative standard deviation less than 3%.Test
For item to substrate METHOD FOR CONTINUOUS DETERMINATION 10 times, test relative deviation is 2.8%.
Above-mentioned test result shows sensor sensitivity with higher, faster response time and good consistency
And repeatability.
Application Example 2
By organophosphorus pesticide biosensor assay organophosphorus pesticide DDVP made from embodiment 2.Temperature is 30 DEG C, bottom
Object (acecoline solution) concentration is 8 × 10-3mol·L-1, in pH=7.0 buffer solution, incubation of the enzyme membrane in pesticide
Time is 4min.Linear sensor response range is 1~10mg/L, and Monitoring lower-cut is up to 0.4mg/L.
Application Example 3
By organophosphorus pesticide biosensor assay organophosphorus pesticide DDVP made from embodiment 3.Temperature is 30 DEG C, bottom
Object (acecoline solution) concentration is 8 × 10-3mol·L-1, in pH=7.0 buffer solution, incubation of the enzyme membrane in pesticide
Time is 6min.Linear sensor response range is 1~10mg/L, and Monitoring lower-cut is up to 0.4mg/L.
Application Example 4
By organophosphorus pesticide biosensor assay organophosphorus pesticide DDVP made from embodiment 4.Temperature is 30 DEG C, bottom
Object (acecoline solution) concentration is 8 × 10-3mol·L-1, in pH=7.0 buffer solution, incubation of the enzyme membrane in pesticide
Time is 3min.Linear sensor response range is 2~8mg/L, and Monitoring lower-cut is up to 0.2mg/L.
Application Example 5
By organophosphorus pesticide biosensor assay organophosphorus pesticide DDVP made from embodiment 5.Temperature is 30 DEG C, bottom
Object (acecoline solution) concentration is 8 × 10-3mol·L-1, in pH=7.0 buffer solution, incubation of the enzyme membrane in pesticide
Time is 3min.Linear sensor response range is 2~6mg/L, and Monitoring lower-cut is up to 0.1mg/L.
Claims (3)
1. a kind of preparation method of electric potential type organophosphorus biosensor, which comprises the following steps:
(1) preparation of Deacetyled konjac glucomannan colloidal sol
The Na for being 0.8~1.2g/L by 0.8~1.2g Deacetyled konjac glucomannan fine powder and 80~120ml concentration2CO3It is molten
After liquid is mixed into translucent colloidal liquid, it is further continued for 15~25min of stirring, it is molten that Deacetyled konjac glucomannan is made
Glue；
(2) preparation of enzyme membrane
Weigh the acetylcholinesterase, 0.8~1.2ml phosphate buffer solution and 0.8~1.2ml deacetylation evil spirit of 0.8~1.2mg
Taro Glucomannan colloidal sol is uniformly mixed, obtains mixed solution；Take 2~5 μ L mixing drop-coated in polyethylene terephthalate
In organic nucleopore membranes, then it is placed in refrigerator freeze-drying lyophilization at a temperature of 4 DEG C, obtains enzyme membrane；
(3) preparation of electric potential type organophosphorus biosensor
Enzyme membrane is fixed on the combination electrode of pH meter with ' O ' shape circle, obtains electric potential type organophosphorus biosensor.
2. preparation method according to claim 1, which is characterized in that in step (2), polyethylene terephthalate has
The aperture of machine nucleopore membranes is 1-10 μm.
3. it is a kind of according to claim 1~one of 2 described in electric potential type organophosphorus biosensor made from preparation method.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050455A (en) * | 2007-03-29 | 2007-10-10 | 中国农业科学院农产品加工研究所 | Immobilization lipase, and preparation method |
| CN104198559A (en) * | 2014-09-15 | 2014-12-10 | 常熟理工学院 | Electrochemical biosensor for organic phosphorus pesticide detection and preparation method of electrochemical biosensor |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IL153012A0 (en) * | 2002-11-21 | 2003-06-24 | Yissum Res Dev Co | Method and device for detecting analytes |
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2018
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101050455A (en) * | 2007-03-29 | 2007-10-10 | 中国农业科学院农产品加工研究所 | Immobilization lipase, and preparation method |
| CN104198559A (en) * | 2014-09-15 | 2014-12-10 | 常熟理工学院 | Electrochemical biosensor for organic phosphorus pesticide detection and preparation method of electrochemical biosensor |
Non-Patent Citations (3)
| Title |
|---|
| 基于脱乙酰基魔芋葡甘聚糖固定酶的H2O2生物传感器;郭晓明等;《分析测试学报》;20080630;第27卷(第6期);第581-584页 * |
| 基于脱乙酰基魔芋葡甘聚糖固定酶的葡萄糖传感器;郭晓明等;《分析试验室》;20071031;第26卷(第10期);第100-103页 * |
| 快速检测有机磷及氨基甲酸酯类农药的酶生物传感器研究;田园;《中国优秀硕士学位论文全文数据库 信息科技辑》;20070115(第01期);第1-67页 * |
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