CN104007154B - A kind of preparation method and application of the redox hydrosol, the integrated microorganism electrode quickly detected for aqueous bio toxicity - Google Patents
A kind of preparation method and application of the redox hydrosol, the integrated microorganism electrode quickly detected for aqueous bio toxicity Download PDFInfo
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Abstract
The present invention relates to a kind of redox hydrosol, prepare with the following method：Organic-inorganic composite sol is prepared with polymeric sol and amino silicane coupling agent, this complex sol and electron mediator are mixed with the redox hydrosol.The invention further relates to a kind of preparation method and application of the integrated microorganism electrode quickly detected for aqueous bio toxicity, pass through the prepared redox hydrosol, microorganism and electron mediator are also secured on electrode, integrated microorganism electrode is prepared, and integrated microorganism electrode is applied in the quick detection of aqueous bio toxicity and at-once monitor.The electrochemical sensor system built using the integrated microorganism electrode prepared by the present invention, can at-once monitor aqueous bio toxicity change and quick detection aqueous bio toxicity size, reach instant, continuous, quick detection, have the characteristics that sensitivity for analysis is high, of low cost, easy to operate, easy to carry.
Description
Technical field
The invention belongs to microbiological sensor technical field, and in particular to the oxidation quickly detected for aqueous bio toxicity
Reduce the hydrosol, the preparation method and application of integrated microorganism electrode.
Background technology
With the development of the modern industry, increasing environmental pollution causes very big impact to aquatic ecosystem, right
It carries out the important step that toxicity detection has become evaluation quality of water environment.It is presently used for the method for pollutant toxotest
Main rationalization method and biological method.Traditional physico-chemical method can be analyzed a certain or certain a class of pollutant with accurate quantification
Species and content；But these methods cannot directly, comprehensively reflect combined influence of the noxious material to environment.Biological method is
By detecting change of the toxicant to biological physiology behavior, and then reflect water body toxicity size, it can more fully reflect useless
The joint toxic action of combined pollutant in water, and it is right to be fully understood by various envirment factors (such as pH value, temperature, solubility)
The specific influence of pollutant poisonous effect, there is very big advantage.Therefore, in water pollution research, as conventional physico-chemical method
Effectively supplement, has become monitoring using biological method progress bio-toxicity detection and evaluates the important hand of water body environment quality
One of section.
Mediator type microbiological sensor had been risen in recent years applies research in bio-toxicity detection field.This method tool
Have that cost is low, it is easy to operate, detect the features such as sensitive.It not only overcomes in national standard method luminescence method easily by the shadow of system turbidity
Ring, BOD method（BOD）Middle dissolved oxygen concentration is low, it is unstable the shortcomings of.And with portable, sensitive, quick and can be real
When on-line analysis the features such as detecting, be expected to be used for the composite acuity bio-toxicity continuous on-line detections of a large amount of polluted-waters.
For mediator type sensor, electron mediator, which is substantially, to be disperseed in the solution.Patent No. 200810036317.5
Chinese patent《The microorganism electrode preparation method of water quality toxicity biosensor》Disclose it is a kind of prepared using double faced adhesive tape it is micro-
The method of biological sensor electrode, the method production electrode on only include microorganism species, in the detection process of sensor
Still need to rely on scattered electron mediator in the solution, and to disperse consumption in the solution larger for electron mediator, unavoidably
On detection sample or reference electrode have certain pollution and influence.
The content of the invention
The invention solves first technical problem be to prepare a kind of redox hydrosol, this colloidal sol has overcome
The shortcomings that machine material swellability and inorganic material easy embrittlement, the advantages of combining both, and electron mediator has been fixed on molten
In glue, preparation method is simple, and bio-compatibility is good.
The invention solves second technical problem be to provide a kind of one quickly detected for aqueous bio toxicity
Change the preparation method of microorganism electrode, this method uses the redox hydrosol prepared by organic-inorganic composite sol by micro- life
Thing flora and electron mediator are fixed on electrode plate, simplify the operating procedure of electrode, while reduce the consumption of electron mediator
Amount, reduces pollution of the electron mediator to detection sample and reference electrode, and is easy to instrumentation and the scene of electrochemical sensor
Detection, is easy to miniaturization, the portability of detection device, improves the convenience and speed of detection, can be used for the detection of no reagent,
And disposable electrode use can be made.
The invention solves the 3rd technical problem be the integrated microorganism that is quickly detected for aqueous bio toxicity
The application of electrode, at-once monitor and prison are used to prepare using the integrated microorganism electrode of redox hydrosol fixation of microbe
The system for surveying aqueous bio toxicity, which includes working electrode, to electrode, reference electrode, electrolytic cell, in at-once monitor water body
During the change of bio-toxicity and detection aqueous bio toxicity size, reach instant, online, continuous detection, not only may be used
To realize quick detection, and have the characteristics that sensitivity for analysis is high, of low cost, easy to operate, easy to carry.
In order to solve first technical problem, the present invention prepares the redox hydrosol with the following method：Will be natural high
Molecular material is dissolved in water preparation organic solution, adjusts pH to 3 ~ 7, adds amino silicane coupling agent, is stirred at 30 ~ 70 DEG C
To the uniform Inorganic whisker colloidal sol of formation；Inorganic whisker colloidal sol is taken to be mixed with electron mediator aqueous solution, when stirring 2 is small
More than, form the redox hydrosol.
Preferably, the high molecular material is gelatin or chitosan；Natural macromolecular material in prepared organic solution
Mass concentration be 10mg/ml ~ 100mg/ml.
Preferably, the amino silicane coupling agent is selected from aminopropyl trimethoxysilane（APTMOS）, three ethoxy of aminopropyl
Base silane（APTEOS）Or N- (β-aminoethyl)-γ-aminopropyltrimethoxysilane；Preferably, amino in the complex sol
The mass fraction of silane coupling agent is 0.5% ~ 5%.
Preferably, the electron mediator includes benzoquinones, 2,6- phlorones or menadione；Preferably, the electronics is situated between
Body concentration of aqueous solution is 0.01mol/L ~ 0.1mol/L；The body that the Inorganic whisker colloidal sol is mixed with electron mediator aqueous solution
Product is than being 10:1~1:10.
In order to solve second technical problem, the present invention is prepared with the following method quickly to be detected for aqueous bio toxicity
Integrated microorganism electrode, specifically comprise the following steps：
（1）Cultivate microbial inoculum；
（2）Take the redox hydrosol to be uniformly mixed with microbial inoculum, obtain mixed sols；
（3）Mixed sols drop coating is taken in electrode surface, dry, cleaning, dries, obtain the redox hydrosol and fix again
The integrated microorganism electrode of microorganism.
Preferably, the microbial inoculum is the mixed of Escherichia coli bacteria liquid, saccharomycete bacterium solution or E.coli and yeast
Close bacterium solution；Preferably, the volume ratio that the redox hydrosol is mixed with microbial inoculum is 1:3~3:1；Preferably, it is described
Electrode includes gold electrode, platinum electrode, glass-carbon electrode or boron doped diamond film electrode.
Preferably, the mixed proportion of E.coli and yeast is 40 ~ 100 parts of weights of Escherichia coli in the mixed bacteria liquid
20 ~ 80 parts of amount, saccharomycete weight.
In order to solve the 3rd technical problem, prepared integrated microorganism electrode is applied to aqueous bio by the present invention
Toxicity quickly detects.
Quickly the application in detection, its concrete application are as follows in aqueous bio toxicity for the integration microorganism electrode：
In three-electrode system, working electrode is the integrated microorganism electrode of the redox hydrosol fixation of microbe prepared, in perseverance
During potentiometer under current-mode, bacterial respiratory nutritive solution is added, sets operating voltage, after background current stabilization, addition is treated
Detect liquid, the change of record current, according to electric current in electric current suppression curve and inhibiting rate relational expression：Inhibiting rate %=(1-I2/
I1) × 100%, I1To add the steady-state current before solution to be detected；I2To add the steady-state current after solution to be detected, suppression is calculated
Rate processed；By the relation between inhibiting rate and solution bio-toxicity to be detected, carry out at-once monitor and detect the biology poison of water body
Property.
Preferably, the operating voltage set is 0.2V ~ 0.7V.
The present invention has the advantages that：
1. the fixation material for fixation of microbe component influences very big, prepared by present invention oxidation for sensor performance
The shortcomings that reducing the hydrosol, overcoming organic material swellability and inorganic material easy embrittlement, the advantages of combining both, and handle
Electron mediator has been fixed in colloidal sol, and preparation method is simple, and bio-compatibility is good.
2. by the redox hydrosol, electron mediator and microorganism can be fixed on the electrode at the same time, prepared
Integrated microorganism electrode.During integrated microorganism electrode use, eliminate and electron mediator is added into solution and is waited
The step that electric current is stablized, shortens detection time, while also reduces the consumption of electron mediator, is so avoided that electron mediator
Pollution to detection sample or reference electrode, and it is easy to the instrumentation and Site Detection of electrochemical sensor, it is easy to detection and sets
Standby miniaturization, portability, improve the convenience and speed of detection, and available for the detection of no reagent, and can be made disposable
Electrode uses.
3. water body life prepared by the integrated microorganism electrode of redox hydrosol fixation of microbe using the present invention
Thing Toxicity Monitoring and monitoring system, change and detection aqueous bio toxicity size available for at-once monitor aqueous bio toxicity,
Reach instant, online, continuous detection, not only realize quick detection, and with sensitivity for analysis is high, of low cost, operation
Simply, the features such as easy to carry.
Brief description of the drawings
The attached drawing that the present invention is included is used to provide a further understanding of the present invention.
Fig. 1 is the detection various concentrations Hg of example 12+Chronoa mperometric plot
Fig. 2 is the Hg of example 12+Concentration-suppression rate response curve
Fig. 3 is the heavy metallic mixture toxicity inhibition rate figure of example 2
Fig. 4 is the DCP of example 52+Concentration-suppression rate response curve
Fig. 5 is the Ni of example 62+Concentration-suppression rate response curve
Embodiment
In order to be better understood from the present invention, with reference to embodiment, the present invention is described further, but the present invention
Content is not limited solely to the following examples.
Embodiment 1：
1st, a kind of preparation of the redox hydrosol：
The mass concentration that Gelatin is obtained in water is the solution of 50mg/ml；PH to 4 is adjusted, adds aminopropyl three
Methoxy silane（APTMOS）, the mass fraction of APTMOS is 1% in solution, and stirring is uniform compound molten to being formed at 50 DEG C
Glue；Complex sol is taken to be mixed with 0.04mol/L benzoquinones colloidal sols（Volume ratio is 4:1）, when stirring 12 is small, to formation redox water
Colloidal sol；
2nd, the preparation of the integrated microorganism electrode quickly detected for aqueous bio toxicity：
（1）Microbial inoculum：With Escherichia coli（E.coli）For challenge organisms, it is inoculated in fluid nutrient medium.Large intestine
Bacillus Liquid Culture based component（Mass fraction）：0.3% beef extract, 1% peptone, 0.5% sodium chloride.Cultivate 16 it is small when, centrifugation is clear
Wash to obtain wet thallus, be dispersed in phosphate buffer, be stored at 4 DEG C；
（2）The redox hydrosol of synthesis is taken, with Escherichia coli bacteria liquid with volume ratio 1:1 ratio is uniformly mixed, and is obtained
To the redox hydrosol and the mixed sols of Escherichia coli；
（3）Mixed sols drop coating is taken to be dried at room temperature in electrode surface；Then cleaned twice with phosphate buffer,
Drying at room temperature, obtains the integrated electrode of redox hydrosol fixation of microbe.
3rd, integrated microorganism electrode is applied to the at-once monitor of aqueous bio toxicity and quick detection：
One integrated microorganism electrode is fixed on a bottom by a stainless steel shelf the electrolysis of circular aperture
The bottom outer in pond, a stainless steel plate is connected in the another side of the electrode；The integrated electrode and the electrolytic cell
Between use rubber seal（The outside diameter of rubber seal is not more than the length of side of working electrode, gasket diameters 3mm）Sealed；
Among the electrolytic cell is placed in as the platinum electrode to electrode and as silver/silver chloride electrode of reference electrode, with described one
Body microorganism electrode is as working electrode；After electrochemical detection device is completed, be followed successively by from down to up stainless steel plate,
Working electrode, rubber seal, electrolytic cell.
Choose pH=7.0, containing 10mmol/L sodium lactates, 10mmol/L sodium succinates, 10mmol/L glucose
0.01mol/L phosphate buffers are bacterial respiratory nutritive solution.
10ml bacterial respiratory nutritive solutions are added into the electrolytic cell of the electrochemical detection device of the three-electrode system.
Then curent change is measured under 0.5V (vs.Ag/AgCl).After about 600s or so current signal stables, various concentrations are added
Solution to be measured carry out toxotest.This process is recorded the change curve of the microbiological sensor electric current by electrochemical workstation,
According to electric current suppression curve, the corresponding inhibiting rate size of various concentrations gradient can be calculated.Inhibiting rate and inhibiting substances concentration it
Between relational expression be：Inhibiting rate %=(1-I2/I1)×100%
Configure the Hg of various concentrations2+Solution carries out water quality acute biological toxicity test as solution to be measured, and chrono-amperometric is bent
Line is as shown in Figure 1, the results show, it can be seen that adds Hg2+20 seconds homeostasis electric currents reduce rapidly afterwards, show the integration prepared
Microorganism electrode can show the mutation of water body Poisoning material concentration in a short period of time, the electric current after noxious material is added
Have and decline to some extent, this shows heavy metal ion Hg2+There is apparent poisonous effect to Escherichia coli.Pass through concentration-suppression
Rate response curve Fig. 2 processed, with interpolation method, obtains Hg2+503nhibiting concentration IC50For 21.2 μ g/ml.It can be seen from the above that the micro- life of integration
Thing electrode can realize the at-once monitor of aqueous bio toxicity and quick detection.
Embodiment 2
Operating method and step are with embodiment 1, unique change：In application continuum microorganism electrode to aqueous bio
During toxicity is monitored and detects, Cu is added into system2+And Zn2+Mixture, Cu2+And Cd2+Mixture, Zn2 +And Cd2+Mixture with instead of Hg2+Solution（Concentration in mixture per heavy metal species is all 40 μ g/ml）.The results show that
Response current is decreased obviously after adding different binary heavy metallic mixtures, and three groups of binary heavy metal mixing are obtained by calculation
The inhibiting rate of thing is as shown in figure 3, toxicity size order is Cu2+-Zn2+>Zn2+-Cd2+>Cu2+-Cd2+
Embodiment 3
1st, a kind of preparation of the redox hydrosol：
The mass concentration that Gelatin is obtained in water is the solution of 10mg/ml；PH to 3 is adjusted, adds aminopropyl three
Methoxy silane（APTMOS）, APTMOS mass fractions are 0.5% in solution, and stirring is uniform compound molten to being formed at 60 DEG C
Glue；Concentration is taken to be mixed for 0.01mol/L menadiones aqueous solution with the complex sol prepared（Volume ratio is 1:10）, it is small to stir 12
When, to the formation redox hydrosol.
2nd, the integrated microorganism electrode quickly detected for aqueous bio toxicity and its preparation：
（1）Microbial inoculum：With Escherichia coli（E.coli）For challenge organisms, it is inoculated in fluid nutrient medium.Large intestine
Bacillus Liquid Culture based component（Mass fraction）：0.3% beef extract, 1% peptone, 0.5% sodium chloride.Cultivate 16 it is small when, centrifugation is clear
Wash to obtain wet thallus, be dispersed in phosphate buffer, be stored at 4 DEG C
（2）The redox hydrosol of synthesis is taken, with Escherichia coli bacteria liquid with volume ratio 1:3 ratio is uniformly mixed, and is obtained
To the redox hydrosol and the mixed sols of Escherichia coli.
（3）Mixed sols drop coating is taken to be dried at room temperature in electrode surface；Then cleaned twice with phosphate buffer,
Drying at room temperature, obtains the integrated electrode of redox hydrosol fixation of microbe.
3rd, integrated microorganism electrode is applied to the at-once monitor of aqueous bio toxicity and quick detection：
One integrated microorganism electrode is fixed on a bottom by a stainless steel shelf the electrolysis of circular aperture
The bottom outer in pond, a stainless steel plate is connected in the another side of the electrode；The integrated electrode and the electrolytic cell
Between use rubber seal（The outside diameter of rubber seal is not more than the length of side of working electrode, gasket diameters 3mm）Sealed；
Among the electrolytic cell is placed in as the platinum electrode to electrode and as silver/silver chloride electrode of reference electrode, with described one
Body microorganism electrode is as working electrode；After electrochemical detection device is completed, be followed successively by from down to up stainless steel plate,
Working electrode, rubber seal, electrolytic cell.
Choose pH=7.0, containing 10mmol/L sodium lactates, 10mmol/L sodium succinates, 10mmol/L glucose
0.01mol/L phosphate buffers are bacterial respiratory nutritive solution.
10ml bacterial respiratory nutritive solutions are added into the electrolytic cell of the electrochemical detection device of the three-electrode system.
Then curent change is measured under 0.2V (vs.Ag/AgCl).After about 600s or so current signal stables, various concentrations are added
Sample to be tested carries out toxotest.This process is recorded the change curve of the microbiological sensor electric current, root by electrochemical workstation
According to electric current suppression curve, the corresponding inhibiting rate size of various concentrations gradient can be calculated.Between inhibiting rate and inhibiting substances concentration
Relational expression is：Inhibiting rate %=(1-I2/I1)×100%
Configure the Cu of 80 μ g/ml2+Water quality acute biological toxicity test is carried out as detected solution, measures the Cu of 80 μ g/ml2 +Inhibiting rate be 37.1%.
Embodiment 4
Operating method and step are with embodiment 3 with the difference of embodiment 3：Preparing the redox hydrosol
During, the electron mediator aqueous solution of addition is 0.02mol/L2,6- phlorones.
Configure the Cu of 80 μ g/ml2+Water quality acute biological toxicity test is carried out as detected solution, measures the Cu of 80 μ g/ml2 +Inhibiting rate be 28.9%.
Embodiment 5
1st, a kind of preparation of the redox hydrosol：
Dissolve chitosan in the solution that the mass concentration obtained in water is 10mg/ml；PH to 7 is adjusted, adds aminopropyl
Trimethoxy silane（APTMOS）, APTMOS mass fractions are 5% in solution, and stirring is uniform compound molten to being formed at 60 DEG C
Glue；Concentration is taken to be mixed for 0.1mol/L benzoquinones aqueous solution with the complex sol prepared（Volume ratio is 1:10）, when stirring 12 is small, extremely
Form the redox hydrosol.
2nd, the integrated microorganism electrode quickly detected for aqueous bio toxicity and its preparation：
（1）Microbial inoculum：Using saccharomycete as challenge organisms, saccharomyces cerevisiae is inoculated with（S.cerevisiae）Trained in liquid
Support in base, Liquid Culture based component is（Mass concentration）：1% yeast extract, 2% peptone, 2% glucose, grow 24h, eccentric cleaning
Wet thallus is obtained, is dispersed in phosphate buffer, is stored at 4 DEG C.
（2）The redox hydrosol of synthesis is taken, with saccharomycete bacterium solution with volume ratio 1:3 ratio is uniformly mixed, and is obtained
The mixed sols of the redox hydrosol and saccharomycete.
（3）Mixed sols drop coating is taken to be dried at room temperature in electrode surface；Then cleaned twice with phosphate buffer,
Drying at room temperature, obtains the integrated electrode of redox hydrosol fixation of microbe.
3rd, integrated microorganism electrode is applied to the at-once monitor of aqueous bio toxicity and quick detection：
One integrated microorganism electrode is fixed on a bottom by a stainless steel shelf the electrolysis of circular aperture
The bottom outer in pond, a stainless steel plate is connected in the another side of the electrode；The integrated electrode and the electrolytic cell
Between use rubber seal（The outside diameter of rubber seal is not more than the length of side of working electrode, gasket diameters 3mm）Sealed；
Among the electrolytic cell is placed in as the platinum electrode to electrode and as silver/silver chloride electrode of reference electrode, with described one
Body microorganism electrode is as working electrode；After electrochemical detection device is completed, be followed successively by from down to up stainless steel plate,
Working electrode, rubber seal, electrolytic cell.
Choose pH=7.0, containing 10mmol/L sodium lactates, 10mmol/L sodium succinates, 10mmol/L glucose
0.01mol/L phosphate buffers are bacterial respiratory nutritive solution.
10ml bacterial respiratory nutritive solutions are added into the electrolytic cell of the electrochemical detection device of the three-electrode system.
Then curent change is measured under 0.7V (vs.Ag/AgCl).After about 600s or so current signal stables, various concentrations are added
Sample to be tested carries out toxotest.This process is recorded the change curve of the microbiological sensor electric current, root by electrochemical workstation
According to electric current suppression curve, the corresponding inhibiting rate size of various concentrations gradient can be calculated.Between inhibiting rate and inhibiting substances concentration
Relational expression is：Inhibiting rate %=(1-I2/I1)×100%
Configure the 3,5- chlorophenesic acids of various concentrations（DCP）Water quality acute biological toxicity test is carried out as detected solution,
As shown in figure 4, obtaining DCP concentration-suppression rate response curve by the inhibiting rate result tested, by interpolation method, obtain DCP half and press down
Concentration IC processed50For 54.3 μ g/ml.
Embodiment 6
1st, a kind of preparation of the redox hydrosol：
The mass concentration that Gelatin is obtained in water is the solution of 100mg/ml；PH to 5 is adjusted, adds aminopropyl three
Ethoxysilane（APTEOS）, the mass fraction of APTEOS is 3% in solution, and stirring is uniform compound molten to being formed at 50 DEG C
Glue；Concentration is taken to be mixed for 0.01mol/L benzoquinones aqueous solution with the complex sol prepared（Volume ratio is 10:1）, when stirring 12 is small,
To the formation redox hydrosol.
2nd, the integrated microorganism electrode quickly detected for aqueous bio toxicity and its preparation：
（1）Microbial inoculum：With Escherichia coli（E.coli）And saccharomyces cerevisiae（S.cerevisiae）Hybrid bacterial strain make
For challenge organisms, the Escherichia coli of freeze-drying are taken（50 parts）And saccharomyces cerevisiae（20 parts）Carry out bacterial strain mixing compounding.Take multiple
The hybrid bacterial strain prepared is added in phosphate buffer solution, is stored at 4 DEG C.
（2）The redox hydrosol of synthesis is taken, with hybrid bacterial strain bacterium solution with volume ratio 3:1 ratio is uniformly mixed, and is obtained
To the redox hydrosol and the mixed sols of hybrid bacterial strain.
（3）Mixed sols drop coating is taken to be dried at room temperature in electrode surface；Then cleaned twice with phosphate buffer,
Drying at room temperature, obtains the integrated electrode of redox hydrosol fixation of microbe.
3rd, it is applied to the instant of aqueous bio toxicity for the integrated microorganism electrode that aqueous bio toxicity quickly detects
Monitoring and quick detection：
One integrated microorganism electrode is fixed on a bottom by a stainless steel shelf the electrolysis of circular aperture
The bottom outer in pond, a stainless steel plate is connected in the another side of the electrode；The integrated electrode and the electrolytic cell
Between use rubber seal（The outside diameter of rubber seal is not more than the length of side of working electrode, gasket diameters 3mm）Sealed；
Among the electrolytic cell is placed in as the platinum electrode to electrode and as silver/silver chloride electrode of reference electrode, with described one
Body microorganism electrode is as working electrode；After electrochemical detection device is completed, be followed successively by from down to up stainless steel plate,
Working electrode, rubber seal, electrolytic cell.
Choose pH=7.0, containing 10mmol/L sodium lactates, 10mmol/L sodium succinates, 10mmol/L glucose
0.01mol/L phosphate buffers are bacterial respiratory nutritive solution.
10ml bacterial respiratory nutritive solutions are added into the electrolytic cell of the electrochemical detection device of the three-electrode system.
Then curent change is measured under 0.3V (vs.Ag/AgCl).After about 600s or so current signal stables, various concentrations are added
Sample to be tested carries out toxotest.This process is recorded the change curve of the microbiological sensor electric current, root by electrochemical workstation
According to electric current suppression curve, the corresponding inhibiting rate size of various concentrations gradient can be calculated.Between inhibiting rate and inhibiting substances concentration
Relational expression is：Inhibiting rate %=(1-I2/I1)×100%
Configure the Ni of various concentrations2+Solution carries out water quality acute biological toxicity test as detected solution, as shown in figure 5,
Ni is obtained by the inhibiting rate result tested2+Concentration-suppression rate response curve, by interpolation method, obtains Ni2+503nhibiting concentration IC50
For 86.2 μ g/ml.
Obviously, the above embodiment of the present invention is only intended to clearly illustrate example of the present invention, and is not pair
The restriction of embodiments of the present invention.For those of ordinary skill in the field, may be used also on the basis of the above description
To make other variations or changes in different ways.Here all embodiments can not be exhaustive.It is every to belong to this hair
Row of the obvious changes or variations that bright technical solution is extended out still in protection scope of the present invention.
Claims (13)
1. a kind of redox hydrosol, it is characterised in that prepared by following methods：Natural macromolecular material is dissolved in water
Middle preparation organic solution, adjusts pH to 3~7, adds amino silicane coupling agent, is stirred at 30~70 DEG C to forming uniform nothing
Machine organic composite colloidal sol；Take Inorganic whisker colloidal sol to be mixed with electron mediator aqueous solution, stirring 2 it is small when more than, formed oxidation
Reduce the hydrosol；
The electron mediator includes benzoquinones, 2,6- phlorones or menadione.
2. the redox hydrosol as claimed in claim 1, it is characterised in that the high molecular material gathers for gelatin or shell
Sugar；The mass concentration of natural macromolecular material is 10mg/ml~100mg/ml in prepared organic solution.
3. the redox hydrosol as claimed in claim 1, it is characterised in that the amino silicane coupling agent is selected from aminopropyl
Trimethoxy silane (APTMOS), aminopropyl triethoxysilane (APTEOS) or N- (β-aminoethyl)-γ-aminopropyl trimethoxy
Base silane.
4. the redox hydrosol as claimed in claim 1, it is characterised in that amino silicone in the Inorganic whisker colloidal sol
The mass fraction of alkane coupling agent is 0.5%~5%.
5. the redox hydrosol as claimed in claim 1, it is characterised in that the electron mediator concentration of aqueous solution is
0.01mol/L~0.1mol/L；The volume ratio that the Inorganic whisker colloidal sol is mixed with electron mediator aqueous solution is 10:1~
1:10。
6. the integrated microorganism for being used for aqueous bio toxicity and quickly detecting is prepared using the redox hydrosol of claim 1
The method of electrode, it is characterised in that include the following steps：
(1) microbial inoculum is cultivated；
(2) take the redox hydrosol to be uniformly mixed with microbial inoculum, obtain mixed sols；
(3) mixed sols drop coating is taken in electrode surface, and dry, cleaning, is dried again, is obtained the redox hydrosol and is fixed micro- life
The integrated microorganism electrode of thing.
7. as claimed in claim 6 prepare the method for being used for the integrated microorganism electrode that aqueous bio toxicity quickly detects,
It is characterized in that, the microbial inoculum is the Mixed Microbes of Escherichia coli bacteria liquid, saccharomycete bacterium solution or E.coli and yeast
Liquid.
8. as claimed in claim 6 prepare the method for being used for the integrated microorganism electrode that aqueous bio toxicity quickly detects,
It is characterized in that, the volume ratio that the redox hydrosol is mixed with microbial inoculum is 1:3~3:1.
9. as claimed in claim 6 prepare the method for being used for the integrated microorganism electrode that aqueous bio toxicity quickly detects,
It is characterized in that, the electrode includes gold electrode, platinum electrode, glass-carbon electrode or boron doped diamond film electrode.
10. as claimed in claim 7 prepare the method for being used for the integrated microorganism electrode that aqueous bio toxicity quickly detects,
It is characterized in that, the mixed proportion of E.coli and yeast is Escherichia coli 40~100 parts of weight, ferment in the mixed bacteria liquid
Female 20~80 parts of weight of bacterium.
11. a kind of utilize such as the integrated microorganism electrode for being used for aqueous bio toxicity and quickly detecting prepared by claim 6
Using, it is characterised in that quickly detected for aqueous bio toxicity.
12. a kind of integrated microorganism electrode quickly detected for aqueous bio toxicity as claimed in claim 11 is answered
With, it is characterised in that in three-electrode system, working electrode is the integration of the redox hydrosol fixation of microbe prepared
Microorganism electrode, under constant voltage chrono-amperometric pattern, adds bacterial respiratory nutritive solution, sets operating voltage, treats background electricity
After stream is stablized, liquid to be detected, the change of record current, according to electric current in electric current suppression curve and inhibiting rate relationship expression are added
Formula：Inhibiting rate %=(1-I2/I1) × 100%, I1To add the steady-state current before solution to be detected；I2It is to be detected molten to add
Steady-state current after liquid, calculates inhibiting rate；By the relation between inhibiting rate and solution bio-toxicity to be detected, supervised immediately
Survey and detect the bio-toxicity of water body.
13. a kind of integrated microorganism electrode quickly detected for aqueous bio toxicity as claimed in claim 12 is answered
With, it is characterised in that the operating voltage set is 0.2V~0.7V.
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| CN112305029A (en) * | 2020-10-23 | 2021-02-02 | 清华大学 | Electrode biological film rapid forming method and microbial electrochemical sensor |
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