CN104007154A - Redox hydrosol, and making method and application of integrated microbial electrode used for rapid detection of biotoxicity of water - Google Patents
Redox hydrosol, and making method and application of integrated microbial electrode used for rapid detection of biotoxicity of water Download PDFInfo
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Abstract
The invention relates to a redox hydrosol. The redox hydrosol is prepared through the steps of preparing an inorganic-organic composite sol by using a polymer sol and an aminosilane coupling agent, and mixing the composite sol with an electronic mediator. The invention also relates to a making method of an integrated microbial electrode used for the rapid detection of the biotoxicity of water, and an application of the integrated microbial electrode. Microbes and the electronic mediator are fixed on an electrode through the prepared redox hydrosol to make the integrated microbial electrode, and the integrated microbial electrode can be applied in the rapid detection and instant monitoring of the biotoxicity of water. An electrochemical sensor can system constructed by using the integrated microbial electrode made in the invention can instantly monitor the water organism toxicity change and rapidly detect the water organism toxicity value, can realize instant, continuous and rapid detection, and has the characteristics of high analysis sensitivity, low cost, simple operation, portability and the like.
Description
Technical field
The invention belongs to microbiological sensor technical field, be specifically related to for the redox hydrosol of water body bio-toxicity fast detecting, preparation method and the application of integrated microorganism electrode.
Background technology
Along with the development of the modern industry, increasing environmental pollution has caused very large impact to aquatic ecosystem, it is carried out to toxicity detection and become the important step of evaluating quality of water environment.At present, the method for pollutant toxotest mainly contains physico-chemical method and biological method.Tradition physico-chemical method can accurate quantification be analyzed kind and the content of a certain or certain first kinds of pollution matter; But these methods can not reflect the combined influence of noxious material to environment directly, all sidedly.Biological method is by detecting the change of toxicant to biological physiology behavior, and then reflection water body toxicity size, it can reflect the joint toxic action of combined pollutant in waste water more all sidedly, and can fully understand the concrete impact of various envirment factors (as pH value, temperature, solubleness etc.) on pollutant poisonous effect, there is very large advantage.Therefore,, in water Pollution Study, as effectively supplementing of conventional physico-chemical method, using biological method to carry out bio-toxicity detection has become monitoring and has evaluated one of important means of water body environment quality.
Rise in recent years mediator type microbiological sensor and be applied in the research of bio-toxicity detection field.It is low that this method has cost, easy to operate, detects the features such as sensitive.The shortcomings such as it has not only overcome the impact that luminescence method in national standard method is subject to system turbidity, and in BOD method (BOD), dissolved oxygen concentration is low, unstable.And have portable, sensitive, quick and can on-line analysis detection etc. feature, be expected to the composite acuity bio-toxicity continuous on-line detection for a large amount of polluted-waters.
For mediator type sensor, electron mediator is to be substantially all dispersed in solution.The patent No. is that the microorganism electrode preparation method > > of 200810036317.5 Chinese patent < < water quality toxicity biosensor discloses a kind of method that adopts double faced adhesive tape to prepare microbiological sensor electrode, on the electrode that the method is produced, only comprise microorganism species, in the testing process of sensor, still need to rely on and be dispersed in the electron mediator in solution, and it is larger that electron mediator is dispersed in solution consumption, inevitably on detecting sample or contrast electrode, there are certain pollution and impact.
Summary of the invention
First technical matters that the present invention will solve is a kind of redox hydrosol of preparation, this colloidal sol has overcome the shortcoming of organic material swellability and the easy embrittlement of inorganic material, combines both advantages, and electron mediator has been fixed in colloidal sol, preparation method is simple, and bio-compatibility is good.
Second technical matters that the present invention will solve is to provide a kind of preparation method of the integrated microorganism electrode for water body bio-toxicity fast detecting, this method adopts the redox hydrosol of being prepared by organic-inorganic composite sol that microorganism species and electron mediator are fixed on battery lead plate, simplified the operation steps of electrode, reduced the consumption of electron mediator simultaneously, reduced electron mediator to detecting the pollution of sample and contrast electrode, and be easy to instrumentation and the Site Detection of electrochemical sensor, be easy to the miniaturization of checkout equipment, portability, improve the convenience and the speed that detect, can be for the detection without reagent, and can make disposable electrode use.
The 3rd technical matters that the present invention will solve is the application for the integrated microorganism electrode of water body bio-toxicity fast detecting, adopt the integrated microorganism electrode of redox hydrosol fixation of microbe for the preparation of the system of at-once monitor and monitoring water body bio-toxicity, this system comprises working electrode, to electrode, contrast electrode, electrolytic cell, in the change of at-once monitor water body bio-toxicity and the process of detection water body bio-toxicity size, reach instant, online, continuous detection, not only can realize fast detecting, and it is high to have sensitivity for analysis, with low cost, simple to operate, the feature such as be easy to carry.
In order to solve first technical matters, the present invention adopts and prepares with the following method the redox hydrosol: natural macromolecular material is dissolved in and in water, prepares organic solution, regulate pH to 3 ~ 7, add amino silicane coupling agent, at 30 ~ 70 ℃, be stirred to and form uniform inorganic organic composite colloidal sol; Get inorganic organic composite colloidal sol and electron mediator aqueous solution, stir more than 2 hours, form the redox hydrosol.
Preferably, described macromolecular material is gelatin or shitosan; In the organic solution of preparing, the mass concentration of natural macromolecular material is 10mg/ml ~ 100mg/ml.
Preferably, described amino silicane coupling agent is selected from aminopropyl trimethoxysilane (APTMOS), aminopropyl triethoxysilane (APTEOS) or N-(β-aminoethyl)-γ-aminopropyltrimethoxysilane; Preferably, in described complex sol, the massfraction of amino silicane coupling agent is 0.5% ~ 5%.
Preferably, described electron mediator comprises benzoquinones, 2,6-phlorone or menadione; Preferably, described electron mediator concentration of aqueous solution is 0.01mol/L ~ 0.1mol/L; The volume ratio of described inorganic organic composite colloidal sol and electron mediator aqueous solution is 10:1 ~ 1:10.
In order to solve second technical matters, the present invention adopts with the following method for the preparation of the integrated microorganism electrode of water body bio-toxicity fast detecting, specifically comprises the steps:
(1) cultivate microbial inoculum;
(2) get the redox hydrosol and mix with microbial inoculum, obtain mixed sols;
(3) get mixed sols and drip and be coated in electrode surface, dry, clean, again dry, obtain the integrated microorganism electrode of redox hydrosol fixation of microbe.
Preferably, described microbial inoculum is the mixed bacteria liquid of Escherichia coli bacteria liquid, saccharomycete bacterium liquid or E.coli and yeast; Preferably, the volume ratio that the described redox hydrosol mixes with microbial inoculum is 1:3 ~ 3:1; Preferably, described electrode comprises gold electrode, platinum electrode, glass-carbon electrode or boron doped diamond film electrode.
Preferably, in described mixed bacteria liquid, the blending ratio of E.coli and yeast is 40 ~ 100 parts of weight of Escherichia coli, 20 ~ 80 parts of weight of saccharomycete.
In order to solve the 3rd technical matters, the present invention is applied to water body bio-toxicity fast detecting by prepared integrated microorganism electrode.
The application of described integrated microorganism electrode in water body bio-toxicity fast detecting, its concrete application is as follows: in three-electrode system, working electrode is the integrated microorganism electrode of the redox hydrosol fixation of microbe of preparation, under constant voltage timing current-mode, add bacterial respiratory nutritive solution, set operating voltage, after background current is stable, add liquid to be detected, the change of record current, suppresses electric current and inhibiting rate relational expression in curve according to electric current: inhibiting rate %=(1-I
2/ I
1) * 100%, I
1for adding the steady-state current before solution to be detected; I
2for adding the steady-state current after solution to be detected, calculate inhibiting rate; By the relation between inhibiting rate and solution bio-toxicity to be detected, carry out the bio-toxicity of at-once monitor and detection water body.
Preferably, the operating voltage of described setting is 0.2V ~ 0.7V.
The present invention has following beneficial effect:
1. very large for sensor performance impact for the immobilization material of fixation of microbe component, the redox hydrosol prepared by the present invention, overcome the shortcoming of organic material swellability and the easy embrittlement of inorganic material, combine both advantages, and electron mediator has been fixed in colloidal sol, preparation method is simple, and bio-compatibility is good.
2. by the redox hydrosol, can electron mediator and microorganism be fixed on electrode, prepare integrated microorganism electrode simultaneously.In integrated microorganism electrode use procedure, saved to adding electron mediator in solution and waiting for the step of current stabilization, shortened detection time, also reduced the consumption of electron mediator simultaneously, can avoid like this electron mediator to detecting the pollution of sample or contrast electrode, and be easy to instrumentation and the Site Detection of electrochemical sensor, be easy to miniaturization, the portability of checkout equipment, improve the convenience and the speed that detect, and can be used for the detection without reagent, and can make disposable electrode use.
3. the monitoring of water body bio-toxicity and the monitoring system that prepared by the integrated microorganism electrode of employing redox hydrosol fixation of microbe of the present invention, can be used for the change of at-once monitor water body bio-toxicity and detect water body bio-toxicity size, reach instant, online, continuous detection, not only realized fast detecting, and had that sensitivity for analysis is high, with low cost, simple to operate, the feature such as be easy to carry.
Accompanying drawing explanation
The accompanying drawing that the present invention comprises is for providing a further understanding of the present invention.
Fig. 1 is the detection variable concentrations Hg of example 1
2+chronoa mperometric plot
Fig. 2 is the Hg of example 1
2+concentration-inhibiting rate response curve
Fig. 3 is the heavy metallic mixture toxicity inhibiting rate figure of example 2
Fig. 4 is the DCP of example 5
2+concentration-inhibiting rate response curve
Fig. 5 is the Ni of example 6
2+concentration-inhibiting rate response curve
Embodiment
For a better understanding of the present invention, below in conjunction with embodiment, the present invention is described further, but content of the present invention is not only confined to the following examples.
Embodiment 1:
1, a kind of preparation of the redox hydrosol:
Gelatin is dissolved in to the solution that the mass concentration that obtains in water is 50mg/ml; Regulate pH to 4, add aminopropyl trimethoxysilane (APTMOS), in solution, the massfraction of APTMOS is 1%, is stirred to and forms uniform complex sol at 50 ℃; Get complex sol and mix (volume ratio is 4:1) with 0.04mol/L benzoquinones colloidal sol, stir 12 hours, to forming the redox hydrosol;
2, for the preparation of the integrated microorganism electrode of water body bio-toxicity fast detecting:
(1) microbial inoculum: take Escherichia coli (E.coli) as tested microorganism, be inoculated in fluid nutrient medium.Escherichia coli Liquid Culture based component (massfraction): 0.3% beef extract, 1% peptone, 0.5% sodium chloride.Cultivate 16 hours, eccentric cleaning obtains wet thallus, is dispersed in phosphate buffer, is kept at 4 ℃;
(2) get the synthetic redox hydrosol, mix with the ratio of Escherichia coli bacteria liquid with volume ratio 1:1, obtain the redox hydrosol and colibacillary mixed sols;
(3) get mixed sols and drip and be coated in electrode surface, at room temperature dry; Then with phosphate buffer, clean twice, drying at room temperature, obtains the integrated electrode of redox hydrosol fixation of microbe.
3, integrated microorganism electrode is applied to at-once monitor and the fast detecting of water body bio-toxicity:
By a stainless steel shelf, an integrated microorganism electrode is fixed on to the bottom outside that the electrolytic cell of circular aperture is arranged at a bottom, at the another side of this electrode, is connected to a stainless steel base plate; Between described integrated electrode and described electrolytic cell, by rubber seal (external diameter of rubber seal is not more than the length of side of working electrode, and packing ring diameter is 3mm), seal; Using as to the platinum electrode of electrode with among the silver/silver chloride electrode of contrast electrode is placed in described electrolytic cell, using described integrated microorganism electrode as working electrode; After electrochemical detection device has been assembled, be followed successively by from down to up stainless steel base plate, working electrode, rubber seal, electrolytic cell.
Choose pH=7.0, contain 10mmol/L sodium lactate, the 0.01mol/L phosphate buffer of 10mmol/L sodium succinate, 10mmol/L glucose is bacterial respiratory nutritive solution.
In the electrolytic cell of the electrochemical detection device of described three-electrode system, add 10ml bacterial respiratory nutritive solution.Then at the lower curent change of measuring of 0.5V (vs.Ag/AgCl).After approximately 600s left and right current signal is stablized, add the solution to be measured of variable concentrations to carry out toxotest.This process is recorded the change curve of this microbiological sensor electric current by electrochemical workstation, according to electric current, suppress curve, can calculate inhibiting rate size corresponding to variable concentrations gradient.Between inhibiting rate and inhibiting substances concentration, relational expression is: inhibiting rate %=(1-I
2/ I
1) * 100%
The Hg of configuration variable concentrations
2+solution carries out the test of water quality acute biological toxicity as solution to be measured, and as shown in Figure 1, result shows, can see and add Hg chronoa mperometric plot
2+within latter 20 seconds, homeostasis electric current reduces rapidly, shows that the integrated microorganism electrode of preparing can show the sudden change of water body Poisoning material concentration in a short period of time, is adding noxious material after-current to have decline to some extent, and this shows heavy metal ion Hg
2+escherichia coli are had to poisonous effect clearly.By concentration-inhibiting rate response curve Fig. 2, by interpolation method, obtain Hg
2+503nhibiting concentration IC
50be 21.2 μ g/ml.As can be seen here, integrated microorganism electrode can be realized at-once monitor and the fast detecting of water body bio-toxicity.
Embodiment 2
Method of operating and step are with embodiment 1, and unique variation is: in the process of water body bio-toxicity being monitored and being detected at application continuum microorganism electrode, in system, add Cu
2+and Zn
2+potpourri, Cu
2+and Cd
2+potpourri, Zn
2+and Cd
2+potpourri to replace Hg
2+solution (in potpourri, the concentration of every heavy metal species is all 40 μ g/ml).Result shows, after adding different binary heavy metallic mixtures, response current has obvious decline, by calculate three groups of binary heavy metallic mixtures inhibiting rate as shown in Figure 3, toxicity size order is Cu
2+-Zn
2+>Zn
2+-Cd
2+>Cu
2+-Cd
2+
Embodiment 3
1, a kind of preparation of the redox hydrosol:
Gelatin is dissolved in to the solution that the mass concentration that obtains in water is 10mg/ml; Regulate pH to 3, add aminopropyl trimethoxysilane (APTMOS), in solution, APTMOS massfraction is 0.5%, is stirred to and forms uniform complex sol at 60 ℃; Getting concentration is that 0.01mol/L menadione aqueous solution is mixed (volume ratio is 1:10) with the complex sol of preparation, stirs 12 hours, to forming the redox hydrosol.
2, for integrated microorganism electrode and the preparation thereof of water body bio-toxicity fast detecting:
(1) microbial inoculum: take Escherichia coli (E.coli) as tested microorganism, be inoculated in fluid nutrient medium.Escherichia coli Liquid Culture based component (massfraction): 0.3% beef extract, 1% peptone, 0.5% sodium chloride.Cultivate 16 hours, eccentric cleaning obtains wet thallus, is dispersed in phosphate buffer, is kept at 4 ℃
(2) get the synthetic redox hydrosol, mix with the ratio of Escherichia coli bacteria liquid with volume ratio 1:3, obtain the redox hydrosol and colibacillary mixed sols.
(3) get mixed sols and drip and be coated in electrode surface, at room temperature dry; Then with phosphate buffer, clean twice, drying at room temperature, obtains the integrated electrode of redox hydrosol fixation of microbe.
3, integrated microorganism electrode is applied to at-once monitor and the fast detecting of water body bio-toxicity:
By a stainless steel shelf, an integrated microorganism electrode is fixed on to the bottom outside that the electrolytic cell of circular aperture is arranged at a bottom, at the another side of this electrode, is connected to a stainless steel base plate; Between described integrated electrode and described electrolytic cell, by rubber seal (external diameter of rubber seal is not more than the length of side of working electrode, and packing ring diameter is 3mm), seal; Using as to the platinum electrode of electrode with among the silver/silver chloride electrode of contrast electrode is placed in described electrolytic cell, using described integrated microorganism electrode as working electrode; After electrochemical detection device has been assembled, be followed successively by from down to up stainless steel base plate, working electrode, rubber seal, electrolytic cell.
Choose pH=7.0, contain 10mmol/L sodium lactate, the 0.01mol/L phosphate buffer of 10mmol/L sodium succinate, 10mmol/L glucose is bacterial respiratory nutritive solution.
In the electrolytic cell of the electrochemical detection device of described three-electrode system, add 10ml bacterial respiratory nutritive solution.Then at the lower curent change of measuring of 0.2V (vs.Ag/AgCl).After approximately 600s left and right current signal is stablized, add variable concentrations testing sample to carry out toxotest.This process is recorded the change curve of this microbiological sensor electric current by electrochemical workstation, according to electric current, suppress curve, can calculate inhibiting rate size corresponding to variable concentrations gradient.Between inhibiting rate and inhibiting substances concentration, relational expression is: inhibiting rate %=(1-I
2/ I
1) * 100%
Configure the Cu of 80 μ g/ml
2+as detected solution, carry out the test of water quality acute biological toxicity, record the Cu of 80 μ g/ml
2+inhibiting rate be 37.1%.
Embodiment 4
Method of operating and step, with embodiment 3, are with the difference of embodiment 3: in preparation redox hydrosol process, the electron mediator aqueous solution adding is 0.02mol/L2,6-phlorone.
Configure the Cu of 80 μ g/ml
2+as detected solution, carry out the test of water quality acute biological toxicity, record the Cu of 80 μ g/ml
2+inhibiting rate be 28.9%.
Embodiment 5
1, a kind of preparation of the redox hydrosol:
Shitosan is dissolved in to the solution that the mass concentration that obtains in water is 10mg/ml; Regulate pH to 7, add aminopropyl trimethoxysilane (APTMOS), in solution, APTMOS massfraction is 5%, is stirred to and forms uniform complex sol at 60 ℃; Getting concentration is that 0.1mol/L benzoquinones aqueous solution is mixed (volume ratio is 1:10) with the complex sol of preparation, stirs 12 hours, to forming the redox hydrosol.
2, for integrated microorganism electrode and the preparation thereof of water body bio-toxicity fast detecting:
(1) microbial inoculum: take saccharomycete as tested microorganism, inoculation saccharomyces cerevisiae (S.cerevisiae) is in fluid nutrient medium, Liquid Culture based component is (mass concentration): 1% yeast extract, 2% peptone, 2% glucose, growth 24h, eccentric cleaning obtains wet thallus, be dispersed in phosphate buffer, be kept at 4 ℃.
(2) get the synthetic redox hydrosol, mix with the ratio of saccharomycete bacterium liquid with volume ratio 1:3, obtain the redox hydrosol and saccharomycetic mixed sols.
(3) get mixed sols and drip and be coated in electrode surface, at room temperature dry; Then with phosphate buffer, clean twice, drying at room temperature, obtains the integrated electrode of redox hydrosol fixation of microbe.
3, integrated microorganism electrode is applied to at-once monitor and the fast detecting of water body bio-toxicity:
By a stainless steel shelf, an integrated microorganism electrode is fixed on to the bottom outside that the electrolytic cell of circular aperture is arranged at a bottom, at the another side of this electrode, is connected to a stainless steel base plate; Between described integrated electrode and described electrolytic cell, by rubber seal (external diameter of rubber seal is not more than the length of side of working electrode, and packing ring diameter is 3mm), seal; Using as to the platinum electrode of electrode with among the silver/silver chloride electrode of contrast electrode is placed in described electrolytic cell, using described integrated microorganism electrode as working electrode; After electrochemical detection device has been assembled, be followed successively by from down to up stainless steel base plate, working electrode, rubber seal, electrolytic cell.
Choose pH=7.0, contain 10mmol/L sodium lactate, the 0.01mol/L phosphate buffer of 10mmol/L sodium succinate, 10mmol/L glucose is bacterial respiratory nutritive solution.
In the electrolytic cell of the electrochemical detection device of described three-electrode system, add 10ml bacterial respiratory nutritive solution.Then at the lower curent change of measuring of 0.7V (vs.Ag/AgCl).After approximately 600s left and right current signal is stablized, add variable concentrations testing sample to carry out toxotest.This process is recorded the change curve of this microbiological sensor electric current by electrochemical workstation, according to electric current, suppress curve, can calculate inhibiting rate size corresponding to variable concentrations gradient.Between inhibiting rate and inhibiting substances concentration, relational expression is: inhibiting rate %=(1-I
2/ I
1) * 100%
3 of configuration variable concentrations, 5-chlorophenesic acid (DCP) carries out the test of water quality acute biological toxicity as detected solution, as shown in Figure 4, by the inhibiting rate result of testing, obtains DCP concentration-inhibiting rate response curve, by interpolation method, obtains DCP 503nhibiting concentration IC
50be 54.3 μ g/ml.
Embodiment 6
1, a kind of preparation of the redox hydrosol:
Gelatin is dissolved in to the solution that the mass concentration that obtains in water is 100mg/ml; Regulate pH to 5, add aminopropyl triethoxysilane (APTEOS), in solution, the massfraction of APTEOS is 3%, is stirred to and forms uniform complex sol at 50 ℃; Getting concentration is that 0.01mol/L benzoquinones aqueous solution is mixed (volume ratio is 10:1) with the complex sol of preparation, stirs 12 hours, to forming the redox hydrosol.
2, for integrated microorganism electrode and the preparation thereof of water body bio-toxicity fast detecting:
(1) microbial inoculum: using the hybrid bacterial strain of Escherichia coli (E.coli) and saccharomyces cerevisiae (S.cerevisiae) as tested microorganism, get cryodesiccated Escherichia coli (50 parts) and saccharomyces cerevisiae (20 parts) and carry out bacterial strain and mix composite.Get composite good hybrid bacterial strain and add in phosphate buffer solution, be kept at 4 ℃.
(2) get the synthetic redox hydrosol, mix with the ratio of hybrid bacterial strain bacterium liquid with volume ratio 3:1, obtain the mixed sols of the redox hydrosol and hybrid bacterial strain.
(3) get mixed sols and drip and be coated in electrode surface, at room temperature dry; Then with phosphate buffer, clean twice, drying at room temperature, obtains the integrated electrode of redox hydrosol fixation of microbe.
3, for the integrated microorganism electrode of water body bio-toxicity fast detecting, be applied to at-once monitor and the fast detecting of water body bio-toxicity:
By a stainless steel shelf, an integrated microorganism electrode is fixed on to the bottom outside that the electrolytic cell of circular aperture is arranged at a bottom, at the another side of this electrode, is connected to a stainless steel base plate; Between described integrated electrode and described electrolytic cell, by rubber seal (external diameter of rubber seal is not more than the length of side of working electrode, and packing ring diameter is 3mm), seal; Using as to the platinum electrode of electrode with among the silver/silver chloride electrode of contrast electrode is placed in described electrolytic cell, using described integrated microorganism electrode as working electrode; After electrochemical detection device has been assembled, be followed successively by from down to up stainless steel base plate, working electrode, rubber seal, electrolytic cell.
Choose pH=7.0, contain 10mmol/L sodium lactate, the 0.01mol/L phosphate buffer of 10mmol/L sodium succinate, 10mmol/L glucose is bacterial respiratory nutritive solution.
In the electrolytic cell of the electrochemical detection device of described three-electrode system, add 10ml bacterial respiratory nutritive solution.Then at the lower curent change of measuring of 0.3V (vs.Ag/AgCl).After approximately 600s left and right current signal is stablized, add variable concentrations testing sample to carry out toxotest.This process is recorded the change curve of this microbiological sensor electric current by electrochemical workstation, according to electric current, suppress curve, can calculate inhibiting rate size corresponding to variable concentrations gradient.Between inhibiting rate and inhibiting substances concentration, relational expression is: inhibiting rate %=(1-I
2/ I
1) * 100%
The Ni of configuration variable concentrations
2+solution carries out the test of water quality acute biological toxicity as detected solution, as shown in Figure 5, by the inhibiting rate result of testing, obtains Ni
2+concentration-inhibiting rate response curve, by interpolation method, obtains Ni
2+503nhibiting concentration IC
50be 86.2 μ g/ml.
Obviously, the above embodiment of the present invention is only for example of the present invention is clearly described, and is not the restriction to embodiments of the present invention.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here cannot give all embodiments exhaustive.Every still row in protection scope of the present invention of apparent variation that technical scheme of the present invention extends out or change that belong to.
Claims (10)
1. the redox hydrosol, it is characterized in that, by following method, prepare: natural macromolecular material is dissolved in water and prepares organic solution, regulate pH to 3 ~ 7, add amino silicane coupling agent, at 30 ~ 70 ℃, be stirred to and form uniform inorganic organic composite colloidal sol; Get inorganic organic composite colloidal sol and electron mediator aqueous solution, stir more than 2 hours, form the redox hydrosol.
2. the redox hydrosol as claimed in claim 1, is characterized in that, preferred, described macromolecular material is gelatin or shitosan; In the organic solution of preparing, the mass concentration of natural macromolecular material is 10mg/ml ~ 100mg/ml.
3. the redox hydrosol as claimed in claim 1, it is characterized in that, preferably, described amino silicane coupling agent is selected from aminopropyl trimethoxysilane (APTMOS), aminopropyl triethoxysilane (APTEOS) or N-(β-aminoethyl)-γ-aminopropyltrimethoxysilane; Preferably, in described inorganic organic composite colloidal sol, the massfraction of amino silicane coupling agent is 0.5% ~ 5%.
4. the redox hydrosol as claimed in claim 1,, it is characterized in that, described electron mediator comprises benzoquinones, 2,6-phlorone or menadione; Preferably, described electron mediator concentration of aqueous solution is 0.01mol/L ~ 0.1mol/L; The volume ratio of described inorganic organic composite colloidal sol and electron mediator aqueous solution is 10:1 ~ 1:10.
5. adopt the redox hydrosol of claim 1 preparation for the preparation of the method for the integrated microorganism electrode of water body bio-toxicity fast detecting, it is characterized in that, comprise the steps:
(1) cultivate microbial inoculum;
(2) get the redox hydrosol and mix with microbial inoculum, obtain mixed sols;
(3) get mixed sols and drip and be coated in electrode surface, dry, clean, again dry, obtain the integrated microorganism electrode of redox hydrosol fixation of microbe.
6. the method for the integrated microorganism electrode for the preparation of water body bio-toxicity fast detecting as claimed in claim 5, it is characterized in that, preferably, described microbial inoculum is the mixed bacteria liquid of Escherichia coli bacteria liquid, saccharomycete bacterium liquid or E.coli and yeast; Preferably, the volume ratio that the described redox hydrosol mixes with microbial inoculum is 1:3 ~ 3:1; Preferably, described electrode comprises gold electrode, platinum electrode, glass-carbon electrode or boron doped diamond film electrode.
7. the method for the integrated microorganism electrode for the preparation of water body bio-toxicity fast detecting as claimed in claim 6, it is characterized in that, preferably, in described mixed bacteria liquid, the blending ratio of E.coli and yeast is 40 ~ 100 parts of weight of Escherichia coli, 20 ~ 80 parts of weight of saccharomycete.
8. utilization, as an application for the integrated microorganism electrode for water body bio-toxicity fast detecting of claim 5 preparation, is characterized in that, for water body bio-toxicity fast detecting.
9. the application of a kind of integrated microorganism electrode for water body bio-toxicity fast detecting as claimed in claim 8, it is characterized in that: in three-electrode system, working electrode is the integrated microorganism electrode of the redox hydrosol fixation of microbe of preparation, under constant voltage timing current-mode, add bacterial respiratory nutritive solution, set operating voltage, after background current is stable, add liquid to be detected, the change of record current, suppresses electric current and inhibiting rate relational expression in curve according to electric current: inhibiting rate %=(1-I
2/ I
1) * 100%, I
1for adding the steady-state current before solution to be detected; I
2for adding the steady-state current after solution to be detected, calculate inhibiting rate; By the relation between inhibiting rate and solution bio-toxicity to be detected, carry out the bio-toxicity of at-once monitor and detection water body.
10. the application of a kind of integrated microorganism electrode for water body bio-toxicity fast detecting as claimed in claim 9, preferred, the operating voltage of described setting is 0.2V ~ 0.7V.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104716336A (en) * | 2015-03-25 | 2015-06-17 | 江西师范大学 | Hydrogel microbial electrode and preparation method thereof |
| CN106770563A (en) * | 2015-11-24 | 2017-05-31 | 中国科学院理化技术研究所 | A kind of bielectron mediator electrochemical biology sensor for detecting water body acute biological toxicity and its application |
| CN112305029A (en) * | 2020-10-23 | 2021-02-02 | 清华大学 | Electrode biological film rapid forming method and microbial electrochemical sensor |
| CN113960141B (en) * | 2021-09-26 | 2023-10-20 | 武汉新烽光电股份有限公司 | Preparation method of immobilized microbial membrane for measuring BOD of seawater |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101261242A (en) * | 2008-04-21 | 2008-09-10 | 同济大学 | Water quality toxicity biosensor microorganism electrode preparation method |
| CN102520047A (en) * | 2011-12-19 | 2012-06-27 | 中国科学院长春应用化学研究所 | Device and method for detecting toxicity of water body |
-
2013
- 2013-02-22 CN CN201310057152.0A patent/CN104007154B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101261242A (en) * | 2008-04-21 | 2008-09-10 | 同济大学 | Water quality toxicity biosensor microorganism electrode preparation method |
| CN102520047A (en) * | 2011-12-19 | 2012-06-27 | 中国科学院长春应用化学研究所 | Device and method for detecting toxicity of water body |
Non-Patent Citations (3)
| Title |
|---|
| HONG WANG ET.AL: "Toxicity assessment of heavy metals and organic compounds using CellSense biosensor with E.coli", 《CHINESE CHEMICAL LETTERS》 * |
| LIHONG LEI ET.AL: "One-pot electrodeposition of 3-aminopropyltriethoxysilane–chitosan hybrid gel film to immobilize glucose oxidase for biosensing", 《SENSORS AND ACTUATORS B: CHEMICAL》 * |
| WENJUAN LI ET.AL: "Immobilization of horseradish peroxidase on chitosan/silica sol–gel hybrid membranes for the preparation of hydrogen peroxide biosensor", 《JOURNAL BIOCHEMICAL BIOPHYSICAL METHODS》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104716336A (en) * | 2015-03-25 | 2015-06-17 | 江西师范大学 | Hydrogel microbial electrode and preparation method thereof |
| CN104716336B (en) * | 2015-03-25 | 2017-11-03 | 江西师范大学 | A kind of hydrogel microorganism electrode and preparation method thereof |
| CN106770563A (en) * | 2015-11-24 | 2017-05-31 | 中国科学院理化技术研究所 | A kind of bielectron mediator electrochemical biology sensor for detecting water body acute biological toxicity and its application |
| CN106770563B (en) * | 2015-11-24 | 2019-04-26 | 中国科学院理化技术研究所 | A kind of bielectron mediator electrochemical biosensor and its application |
| CN112305029A (en) * | 2020-10-23 | 2021-02-02 | 清华大学 | Electrode biological film rapid forming method and microbial electrochemical sensor |
| CN113960141B (en) * | 2021-09-26 | 2023-10-20 | 武汉新烽光电股份有限公司 | Preparation method of immobilized microbial membrane for measuring BOD of seawater |
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| CN104007154B (en) | 2018-04-20 |
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