CN104914152B - The cleaning method of the microelectrode of water body toxicity detection means - Google Patents
The cleaning method of the microelectrode of water body toxicity detection means Download PDFInfo
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Abstract
The invention discloses a kind of cleaning method of the microelectrode of water body toxicity detection means, belong to aquatic monitoring technical field.The microelectrode for solving water body toxicity detection means in the prior art is easily contaminated in detection process, so that the unstable technical problem of the electric current of detection.The method of the present invention is to use more than cleaning fluid cleaning electrode 5min；Wherein, cleaning fluid is water sample to be measured and the mixed solution of microorganism, the concentration OD600 of microorganism is 0.02 0.1 in cleaning fluid, this method can reduce pollution of the test specimens to working electrode in detection process, and then obtain stable current signal output, the deviation of detection electric current even if after multiple test is also typically smaller than 3%, not more than 7%.
Description
Technical field
The invention belongs to aquatic monitoring technical field, and in particular to a kind of cleaning of the microelectrode of water body toxicity detection means
Method.
Background technology
Water body " toxicity " is a brand-new aqueous bio index, directly reflects the contaminated situation of water body.In recent years, I
State there occurs multiple paroxysmal water contamination accident, and life and production and the ecosystem to periphery bring serious shadow
Ring.Therefore, it is necessary to be monitored on-line to the total toxicity of water body.Chinese patent (publication number 102520047A) discloses a kind of inspection
Survey the detection means of water body toxicity and the detection method of water body toxicity.This method utilizes the toxicant in water sample to be measured to micro- life
The inhibitory action of thing breathing, hinders the electron transmission between microorganism and electron mediator, so that by the change of electron transmission,
Judge water body toxicity.The device mainly includes microbial cultivation device and the Electrochemical Detection connected with microbial cultivation device dress
Put composition；Wherein, microbial cultivation device mainly includes filling water sample to be measured, electron mediator and the microculture of microorganism
Ware；Electrochemical detection device mainly includes electrochemical test pond, thermostat and three electrodes being arranged in electrochemical investigating pond
System, three-electrode system is constituted by working electrode, reference electrode and to electrode, reference electrode and to electrode using commonly used in the art
Electrode, but be increase detection sensitivity, working electrode is preferred to use microelectrode or microelectrode array.
In aquatic monitoring technical field, to ensure the stability of testing result, usually require before detection to working electrode
Cleaned, to remove working electrode surface pollutant, regain clean working electrode.Online electrochemical monitoring and analysis
In equipment, the mode of cleaning electrode is mainly physical method, chemical method, method, the photochemical method of physical bond chemistry.
Physical method is mainly rinsed using mechanical cleaning, pulsating water, ultrasonic wave is cleaned, and the pollution of electrode surface is made by physical action
Thing comes off, and reaches the purpose of cleaning electrode；Chemical method is mainly according to contaminated object, using appropriate solution as washing lotion,
The pollutant of electrode surface is set to be dissolved into washing lotion by modes such as immersions；Physics and chemically combined method are mainly in conjunction with thing
The advantage of reason and chemical two methods, chemolysis and mechanical stripping are combined, to reach the purpose of strength cleaning.Photochemistry
Method be that electrode is cleaned using the mode of ultraviolet radiation.But, detection water sample in organic pollution content compared with
In the case of height, in detection process, organic pollution can be adsorbed onto working electrode surface, contaminated electrode, and then cause work electricity
The electric current of pole detection is unstable, and testing result is inaccurate, and especially when working electrode is microelectrode, this influence is stronger.And
All kinds of cleaning methods of the prior art can not all solve this technical problem.
The content of the invention
Present invention aim to address the microelectrode of water body toxicity detection means in the prior art in detection process easily
It is contaminated, so that there is provided a kind of the clear of microelectrode of water body toxicity detection means for the unstable technical problem of the electric current of detection
Washing method.
The technical scheme that the present invention solves above-mentioned technical problem use is as follows.
The cleaning method of the microelectrode of water body toxicity detection means：With more than cleaning fluid cleaning electrode 5min；
The cleaning fluid is that the concentration OD600 of microorganism in water sample to be measured and the mixed solution of microorganism, cleaning fluid is
0.02-0.1。
Further, the water sample to be measured that the cleaning fluid is added after microorganism by diluting is obtained, and the retarder thinner is
Water sample to be measured.
Further, electron mediator is also contained in the mixed solution, further, the cleaning fluid passes through dilution
The water sample to be measured added after electron mediator and microorganism is obtained, and the retarder thinner is water sample to be measured.
Further, the electron mediator is the derivative of the potassium ferricyanide, dimethyl diaminophenazine chloride, ferrocene or ferrocene.
Further, the scavenging period is 5-60min.
Compared with prior art, beneficial effects of the present invention are：
The cleaning method of the microelectrode of the water body toxicity detection means of the present invention, contains electron mediator using after dilution
Water sample to be measured with microorganism is cleaning fluid, under working electrode working condition, makes cleaning fluid continue to connect with working electrode flowing
Touch, the organic matter in cleaning fluid is acted on working electrode surface, pollutant reaches adsorption equilibrium in working electrode surface, form steady
Fixed pollutant adsorption layer, in detection process, because working electrode surface has formd stable pollutant adsorption layer, from
And pollution of the test specimens to working electrode in detection process is reduced, and then stable current signal output is obtained, even if many
The deviation of detection electric current after secondary test is also typically smaller than 3%, not more than 7%.
Brief description of the drawings
Fig. 1 is the ratio of the different carrying currents for detecting the test specimens under number of times and the carrying current of control sample in embodiment 1
Value；
Fig. 2 is the real-time current value and the relation curve of detection time of the detection device records of embodiment 2；
The partial enlarged drawing that Fig. 3 is A in Fig. 2；
Fig. 4 is the real-time current value and the local relation curve of detection time of the detection device records of comparative example 1；
Fig. 5 is the real-time current value and the local relation curve of detection time of the detection device records of comparative example 4；
Fig. 6 is the real-time current value and the local relation curve of detection time of the detection device records of comparative example 5.
Embodiment
In order to further appreciate that the present invention, the preferred embodiments of the invention are retouched with reference to embodiment
State, but it is to be understood that these descriptions are simply to further illustrate the features and advantages of the present invention rather than to patent of the present invention
It is required that limitation.
The cleaning method of the microelectrode of the water body toxicity detection means of the present invention, it is adaptable to containing organic in water sample to be measured
Thing, and working electrode is the detection means of the water body toxicity of microelectrode, it is adaptable to need to add micro- into water sample to be measured when detecting
Biology, and working electrode is the detection means of the water body toxicity of microelectrode, is particularly suitable for use in Chinese patent (publication number
A kind of cleaning for the detection means for detecting water body toxicity disclosed in 102520047A).And the cleaning method is applicable not only to list
Solely detection, also can be used in on-line checking.
The present invention Cleaning principle be：The organic matter contained in water sample to be measured and/or having of being introduced after the microorganism of addition
Machine thing, in detection process, is adsorbed onto working electrode surface, contaminated electrode, influence detection electric current, the present invention makes cleaning fluid first hold
Continuous to be contacted with working electrode flowing, the organic matter in cleaning fluid is acted on working electrode surface, and pollutant is in working electrode surface
Adsorption equilibrium is reached, stable pollutant adsorption layer is formed, in detection process, because working electrode surface has been formd surely
Fixed pollutant adsorption layer, so as to reduce pollution of the test specimens to working electrode in detection process, and then obtains stabilization
Current signal is exported.
The cleaning method of the microelectrode of the water body toxicity detection means of the present invention, is to use cleaning fluid cleaning electrode 5min
More than, i.e., in the state of working electrode work, make cleaning fluid with more than solid stream of water form flushing work electrode 5min, cleaning
Working electrode afterwards is used directly for detecting water sample to be measured, can also be stored in cleaning fluid, during water sample to be measured to be detected, takes
Go out direct use, it is general using first cleaning electrode when for on-line checking, then water sample is detected, electricity is then cleaned again
Pole, detects the continuous circulation working method of water sample again.
Wherein, cleaning fluid is water sample to be measured and the mixed solution of microorganism, and electron mediator does not influence on cleaning process,
So the concentration OD600 that can also also contain microorganism in electron mediator, cleaning fluid in mixed solution is 0.02-0.1.For just
In operation and on-line checking, the water sample to be measured that cleaning fluid typically has been added to electron mediator and microorganism using dilution is obtained,
But retarder thinner is necessary for water sample to be measured, no person is unable to reach the cleaning performance of the present invention.It should be noted that the present invention's is clear
If washing the time less than 5min, organic matter is shorter in the adsorption time of electrode surface, it is impossible to form stable adsorption layer, and then
Stable detection signal can not be obtained.If above 60min, although still can ensure cleaning performance, but be unsatisfactory for instrument
On-line continuous test request, so the preferred 5-60min of scavenging period of the present invention.The concentration OD600 of microorganism is such as in cleaning fluid
Fruit is less than 0.01, then the content of organics in cleaning fluid is too low, it is impossible to which formation can suppress organic suction that electrode further pollutes
Attached layer, and then reliable detection data can not be obtained.If above 0.2, then in During Process of Long-term Operation, easily in electrode surface
The excessive organic matter of accumulation simultaneously forms microbial film, influences the output performance steady in a long-term of microelectrode detector.
In the present invention, the species of the microorganism of addition is not limited, and can be single culture or a variety of strains
Mixing, general microorganism is obtained by water sample Culture in situ to be measured, and Culture in situ method is microorganism in prior art, cleaning fluid
Species and the species of microorganism of the addition in water sample to be measured can be the same or different, it is preferably identical；Electron mediator
For the derivative of the potassium ferricyanide, dimethyl diaminophenazine chloride, ferrocene or ferrocene；The material of microelectrode is the noble metals such as gold, platinum, not special
Limitation.
The present invention is further illustrated with reference to embodiments.
Embodiment 1
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 0.2, and the concentration of the potassium ferricyanide is 2g/L；
Step 3: taking partial test sample, it is diluted with water sample to be measured, is 0.01 to microorganism concn OD600, obtains clear
Washing lotion；
Step 4: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained,
In control sample, microorganism concn OD600 is 0.2, and the concentration of the potassium ferricyanide is 2g/L；
Step 5: microelectrode (20 μm of platinum microelectrodes) 60min of detection means is cleaned with cleaning fluid, then with detection dress
Put and detect control sample and test specimens after microbial cultivation device culture successively, the real-time electricity of micro-current in record detection process
Flow valuve；
Step 6: continuously running detection means, repeat step five is detected 400 times to common.
The water sample to be measured that embodiment 1 is used is deionized water, and detection means is Chinese patent (publication number 102520047A)
The detection means of disclosed detection water body toxicity.Fig. 1 is the limit of the different test specimens detected under number of times of embodiment 1 and control sample
The ratio of electric current, it can be seen that the deviation of the detection electric current after continuous 350 tests is less than 2%, illustrates the present invention
Pollution of the test specimens to working electrode in detection process is reduced, stable current signal output is obtained.
Embodiment 2
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 3: taking partial test sample, it is diluted with water sample to be measured, is 0.01 to microorganism concn OD600, obtains clear
Washing lotion；
Step 4: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained,
In control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 5: microelectrode (25 μm of golden disk microelectrodes) 5min of detection means is cleaned with cleaning fluid, then with detection
Device detects control sample and test specimens after microbial cultivation device culture successively, and micro-current is real-time in record detection process
Current value；
Step 6: continuously running detection means, repeat step five is detected 100 times to common.
The water sample to be measured that embodiment 2 is used is deionized water, and detection means is Chinese patent (publication number 102520047A)
The detection means of disclosed detection water body toxicity.Fig. 2 is the relation song of the real-time current value that embodiment 2 is recorded and detection time
Line, Fig. 3 is Fig. 2 partial enlarged drawing, and as the control sample after cleaning fluid, culture, the test specimens after culture are once continuous successively
Flow through the current signal of microelectrode detector generation, from Fig. 2-3 it can be seen that, although test specimens can be to electrode when flowing through microelectrode
Pollution is produced, but after repeatedly test, the current signal difference of control sample and test specimens is still less than 3%.
Embodiment 3
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 1, and the concentration of the potassium ferricyanide is 3g/L；
Step 3: taking partial test sample, it is diluted with water sample to be measured, is 0.2 to microorganism concn OD600, obtains clear
Washing lotion；
Step 4: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained,
In control sample, microorganism concn OD600 is 1, and the concentration of the potassium ferricyanide is 3g/L；
Step 5: microelectrode (25 μm of golden disk microelectrodes) 60min of detection means is cleaned with cleaning fluid, then with detection
Device detects control sample and test specimens after microbial cultivation device culture successively, and micro-current is real-time in record detection process
Current value；
Step 6: continuously running detection means, repeat step five is detected 120 times to common.
The water sample to be measured that embodiment 3 is used is Songhuajiang River Water, and detection means is Chinese patent (publication number 102520047A)
The detection means of disclosed detection water body toxicity.After tested, the deviation of the detection electric current after continuous 120 tests is less than 2%.
Embodiment 4
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 6, and the concentration of the potassium ferricyanide is 15g/L；
Step 3: taking partial test sample, it is diluted with water sample to be measured, is 0.1 to microorganism concn OD600, obtains clear
Washing lotion；
Step 4: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained,
In control sample, microorganism concn OD600 is 6, and the concentration of the potassium ferricyanide is 15g/L；
Step 5: microelectrode (25 μm of golden disk microelectrodes) 60min of detection means is cleaned with cleaning fluid, then with detection
Device detects control sample and test specimens after microbial cultivation device culture successively, and micro-current is real-time in record detection process
Current value；
Step 6: continuously running detection means, repeat step five is detected 100 times to common.
The water sample to be measured that embodiment 4 is used is Chinese patent (publication number 102520047A) for her logical river, detection means
The detection means of disclosed detection water body toxicity.After tested, the deviation of the detection electric current after continuous 100 tests is less than 2%.
Comparative example 1
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 3: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained,
In control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 4: the potassium ferricyanide using concentration as 2g/L is cleaning fluid, the microelectrode of detection means is first cleaned with cleaning fluid
(20 μm of platinum microelectrodes) 5min, then detects control sample and survey after microbial cultivation device culture successively with detection means
The real-time current value of micro-current in sample, record detection process；
Step 5: continuously running detection means, repeat step four is detected 100 times to common.
The water sample to be measured that comparative example 1 is used is deionized water, and detection means is Chinese patent (publication number 102520047A)
The detection means of disclosed detection water body toxicity.Fig. 4 be comparative example 1 in cleaning fluid, culture after control sample, culture after survey
Sample once continues to flow through microelectrode, the real-time current value of detection device records successively.It can be seen that control sample and survey
The deviation of sample is more than 10%.
Comparative example 2
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 3: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained,
In control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 4: first cleaning detection dress with cleaning fluid with alkaline solution (0.1M KOH are logical containing 0.5% Qula) for cleaning fluid
Microelectrode (20 μm of the platinum microelectrodes) 5min put, is then detected after microbial cultivation device culture successively with detection means
The real-time current value of micro-current in control sample and test specimens, record detection process；
Step 5: continuously running detection means, repeat step four is detected 100 times to common.
The water sample to be measured that comparative example 2 is used is deionized water, and detection means is Chinese patent (publication number 102520047A)
The detection means of disclosed detection water body toxicity.After tested, the deviation of control sample and test specimens is more than 10%.
Comparative example 3
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 3: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained,
In control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 4: first cleaning detection dress with cleaning fluid with acid solution (0.1M KOH are logical containing 0.5% Qula) for cleaning fluid
Microelectrode (20 μm of the platinum microelectrodes) 5min put, is then detected after microbial cultivation device culture successively with detection means
The real-time current value of micro-current in control sample and test specimens, record detection process；
Step 5: continuously running detection means, repeat step four is detected 100 times to common.
The water sample to be measured that comparative example 3 is used is deionized water, and detection means is Chinese patent (publication number 102520047A)
The detection means of disclosed detection water body toxicity.After tested, the deviation of control sample and test specimens is more than 30%.
Comparative example 4
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 3: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained,
In control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 4: using 60 DEG C of deionized water as cleaning fluid, first cleaned with cleaning fluid detection means microelectrode (20 μm
Platinum microelectrode) 5min, then detects control sample and test specimens after microbial cultivation device culture, remembers successively with detection means
Record the real-time current value of micro-current in detection process；
Step 5: continuously running detection means, repeat step four is detected 100 times to common.
The water sample to be measured that comparative example 4 is used is deionized water, and detection means is Chinese patent (publication number 102520047A)
The detection means of disclosed detection water body toxicity.Fig. 5 be comparative example 4 in cleaning fluid, culture after control sample, culture after survey
Sample once continues to flow through microelectrode, the real-time current value of detection device records successively.It can be seen that control sample and survey
The deviation of sample is more than 10%.
Comparative example 5
Step 1: taking water sample to be measured to carry out Culture in situ, microorganism is obtained；
Step 2: taking water sample to be measured again, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, is tested
In sample, test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 3: removing ionized water, the microorganism that the potassium ferricyanide and step one are cultivated is added thereto, control sample is obtained
In, in control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L；
Step 4: using 60 DEG C of deionized water as cleaning fluid, first with cleaning fluid in the form of high voltage pulse (being more than 2kg) it is clear
Microelectrode (20 μm of platinum microelectrodes) 5min of detection means is washed, is then detected successively through microbial cultivation device with detection means
The real-time current value of micro-current in control sample and test specimens after culture, record detection process；
Step 5: continuously running detection means, repeat step four is detected 100 times to common.
The water sample to be measured that comparative example 5 is used is deionized water, and detection means is Chinese patent (publication number 102520047A)
The detection means of disclosed detection water body toxicity.Fig. 6 be comparative example 5 in cleaning fluid, culture after control sample, culture after survey
Sample once continues to flow through microelectrode, the real-time current value of detection device records successively.It can be seen that control sample and survey
The deviation of sample is more than 10%.
It should be noted that to meet single control principle, the water sample phase to be measured that comparative example 1-5 is used with embodiment 2
Together.Comparative example 2 and comparative example 1-5 testing result can be seen that after the cleaning method cleaning electrode using the present invention, drop
Pollution of the test specimens to working electrode in low detection process, results in stable current signal output, even if repeatedly measurement
Deviation also very little.
Obviously, the explanation of above example is only intended to the method and its core concept for helping to understand the present invention.It should refer to
Go out, under the premise without departing from the principles of the invention, can also be to this hair for the those of ordinary skill of the technical field
Bright to carry out some improvement and modification, these are improved and modification is also fallen into the protection domain of the claims in the present invention.
Claims (6)
1. the cleaning method of the microelectrode of water body toxicity detection means, it is characterised in that with cleaning fluid cleaning electrode 5min
More than；
The cleaning fluid is that the concentration OD600 of microorganism in water sample to be measured and the mixed solution of microorganism, cleaning fluid is 0.02-
0.1。
2. the cleaning method of the microelectrode of water body toxicity detection means according to claim 1, it is characterised in that described clear
The water sample to be measured that washing lotion is added after microorganism by diluting is obtained, and the retarder thinner is water sample to be measured.
3. the cleaning method of the microelectrode of water body toxicity detection means according to claim 1, it is characterised in that described clear
Also contain electron mediator in washing lotion.
4. the cleaning method of the microelectrode of water body toxicity detection means according to claim 3, it is characterised in that described clear
The water sample to be measured that washing lotion is added after electron mediator and microorganism by diluting is obtained, and the retarder thinner is water sample to be measured.
5. the cleaning method of the microelectrode of water body toxicity detection means according to claim 3, it is characterised in that the electricity
Sub- mediator is the derivative of the potassium ferricyanide, dimethyl diaminophenazine chloride, ferrocene or ferrocene.
6. the cleaning method of the microelectrode of the water body toxicity detection means according to claim 1-5 any one, its feature
It is, the scavenging period is 5-60min.
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| JP2004223466A (en) * | 2003-01-27 | 2004-08-12 | Yokogawa Electric Corp | Cleaning device |
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| CN102520040A (en) * | 2012-01-09 | 2012-06-27 | 中国科学院长春应用化学研究所 | Microbial reactor for detecting water toxicity and method for detecting water toxicity |
| CN103018306A (en) * | 2012-12-07 | 2013-04-03 | 上海逸臣环保科技有限公司 | Apparatus capable of on-line continuous monitoring of pH value of water sample and automatic cleaning of electrode |
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