CN104914152A - Method for cleaning microelectrode of water toxicity detection device - Google Patents
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Abstract
The invention discloses a method for cleaning a microelectrode of a water toxicity detection device and belongs to the technical field of water monitoring. The method solves the technical problem that based the prior art, a microelectrode in a water toxicity detection device can be polluted easily in detection so that detection current is not stable. The method utilizes a cleaning liquid to clean a working electrode for 5min or more, the cleaning liquid is a mixed solution of a water sample to be detected and microbes, the cleaning liquid has a microbe concentration OD600 of 0.02-0.1, the method reduces pollution produced by a detection sample on the working electrode so that stable current signal output is realized, and after multiple test processes, detection current deviation is generally less than 3% and the highest current deviation is less than or equal to 7%.
Description
Technical field
The invention belongs to aquatic monitoring technical field, be specifically related to a kind of cleaning method of microelectrode of water body toxicity pick-up unit.
Background technology
Water body " toxicity " is a brand-new aqueous bio index, directly reflects the contaminated situation of water body.In recent years, China there occurs repeatedly paroxysmal water contamination accident, brings serious impact to the life of periphery and production and the ecosystem.Therefore, be necessary to carry out on-line monitoring to the total toxicity of water body.Chinese patent (publication number 102520047A) discloses and a kind ofly detects the pick-up unit of water body toxicity and the detection method of water body toxicity.The method utilizes the toxicant in water sample to be measured to the inhibiting effect of microbial respiratory, hinders the electron transmission between microorganism and electron mediator, thus by the change of electron transmission, judges water body toxicity.This device mainly comprises microbial cultivation device and the electrochemical detection device that is communicated with microbial cultivation device forms; Wherein, microbial cultivation device mainly comprises the microbial culture dish filling water sample to be measured, electron mediator and microorganism; The three-electrode system that electrochemical detection device mainly comprises electrochemical test pond, thermostat and is arranged in electrochemical investigating pond, three-electrode system is by working electrode, contrast electrode and form electrode, contrast electrode and this area common electrode is adopted to electrode, but for increasing detection sensitivity, working electrode preferably adopts microelectrode or microelectrode array.
In aquatic monitoring technical field, for ensureing the stability of testing result, usually need before detection to clean working electrode, to remove working electrode surface pollutant, to regain clean working electrode.In online electrochemical monitoring and analytical equipment, the mode of cleaning electrode is mainly physical method, chemical method, the method for physical bond chemistry, photochemical method.Physical method mainly uses the flushing of mechanical cleaning, pulsating water, Ultrasonic Cleaning, by physical action, the pollutant of electrode surface is come off, reaches the object of cleaning electrode; Chemical method, mainly according to contaminated object, adopts suitable solution as washing lotion, makes the pollutant of electrode surface be dissolved in washing lotion by modes such as immersions; Chemical dissolution and mechanical stripping, mainly in conjunction with the advantage of physics and chemistry two kinds of methods, combine by the method that physics and chemistry combines, to reach the object of powerful cleaning.Photochemical method utilizes the mode of ultraviolet radiation to clean electrode.But, in detection water sample, organic contaminant content is higher, in testing process, organic contaminant can be adsorbed onto working electrode surface, contaminated electrode, and then the electric current causing working electrode to detect is unstable, testing result is inaccurate, especially, when working electrode is microelectrode, this impact is stronger.And all kinds of cleaning method of the prior art all can not solve this technical matters.
Summary of the invention
The microelectrode that the object of the invention is to solve water body toxicity pick-up unit in prior art is easily polluted in testing process, so that the technical matters of the electric current instability detected, and provides a kind of cleaning method of microelectrode of water body toxicity pick-up unit.
The technical scheme that the present invention solves the problems of the technologies described above employing is as follows.
The cleaning method of the microelectrode of water body toxicity pick-up unit: with cleaning fluid cleaning more than electrode 5min;
Described cleaning fluid is the mixed solution of water sample to be measured and microorganism, and in cleaning fluid, the concentration OD600 of microorganism is 0.02-0.1.
Further, described cleaning fluid adds the water sample to be measured after microorganism by dilution and obtains, and described diluting solvent is water sample to be measured.
Further, also containing electron mediator in described mixed solution, further, described cleaning fluid adds the water sample to be measured after electron mediator and microorganism by dilution and obtains, and described diluting solvent is water sample to be measured.
Further, described electron mediator is the derivant of the potassium ferricyanide, dimethyl diaminophenazine chloride, ferrocene or ferrocene.
Further, described scavenging period is 5-60min.
Compared with prior art, beneficial effect of the present invention is:
The cleaning method of the microelectrode of water body toxicity pick-up unit of the present invention, the water sample to be measured containing electron mediator and microorganism after dilution is adopted to be cleaning fluid, under working electrode duty, cleaning fluid is made to continue to flow with working electrode to contact, organism in cleaning fluid and working electrode surface effect, pollutant reaches adsorption equilibrium at working electrode surface, form stable pollutant adsorbed layer, in testing process, because working electrode surface has defined stable pollutant adsorbed layer, thus to reduce in testing process test specimens to the pollution of working electrode, and then obtain stable current signal output, even if the deviation of the detection electric current repeatedly after test is also less than 3% usually, the highlyest be no more than 7%.
Accompanying drawing explanation
Fig. 1 is the different ratio detecting the limiting current of test specimens under number of times and the limiting current of control sample in embodiment 1;
Fig. 2 is real-time current value and the relation curve of detection time of the detection device records of embodiment 2;
Fig. 3 is the partial enlarged drawing of A in Fig. 2;
Fig. 4 is the real-time current value of the detection device records of comparative example 1 and the local relation curve of detection time;
Fig. 5 is the real-time current value of the detection device records of comparative example 4 and the local relation curve of detection time;
Fig. 6 is the real-time current value of the detection device records of comparative example 5 and the local relation curve of detection time.
Embodiment
In order to understand the present invention further, below in conjunction with embodiment, the preferred embodiments of the invention are described, but should be appreciated that these describe just as further illustrating the features and advantages of the present invention instead of the restriction to patent requirements of the present invention.
The cleaning method of the microelectrode of water body toxicity pick-up unit of the present invention, be applicable in water sample to be measured containing organism, and working electrode is the pick-up unit of the water body toxicity of microelectrode, need to add microorganism in water sample to be measured when being applicable to detect, and working electrode is the pick-up unit of the water body toxicity of microelectrode, be specially adapted to the disclosed a kind of cleaning detecting the pick-up unit of water body toxicity of Chinese patent (publication number 102520047A).And this cleaning method is not only applicable to independent detection, can be used in on-line checkingi yet.
Cleaning principle of the present invention is: the organism introduced after the organism contained in water sample to be measured and/or the microorganism of interpolation, in testing process, be adsorbed onto working electrode surface, contaminated electrode, impact detects electric current, the present invention makes cleaning fluid first continue to flow with working electrode to contact, organism in cleaning fluid and working electrode surface effect, pollutant reaches adsorption equilibrium at working electrode surface, form stable pollutant adsorbed layer, in testing process, because working electrode surface has defined stable pollutant adsorbed layer, thus to reduce in testing process test specimens to the pollution of working electrode, and then obtain stable current signal output.
The cleaning method of the microelectrode of water body toxicity pick-up unit of the present invention, with cleaning fluid cleaning more than electrode 5min, namely under the state of working electrode work, cleaning fluid is made to rinse more than working electrode 5min with solid stream of water form, working electrode after cleaning can be directly used in and detect water sample to be measured, also can be kept in cleaning fluid, during water sample to be measured to be detected, take out and directly use, when for on-line checkingi, the first cleaning electrode of general employing, then detect water sample, and then cleaning electrode, again detect the continuous circulation working method of water sample.
Wherein, cleaning fluid is the mixed solution of water sample to be measured and microorganism, and electron mediator does not affect cleaning process, so also can also contain electron mediator in mixed solution, in cleaning fluid, the concentration OD600 of microorganism is 0.02-0.1.For ease of operation and on-line checkingi, the water sample to be measured that cleaning fluid generally adopts dilution to add electron mediator and microorganism obtains, but diluting solvent is necessary for water sample to be measured, and no person cannot reach cleaning performance of the present invention.It should be noted that, if scavenging period of the present invention is lower than 5min, then organism is shorter at the adsorption time of electrode surface, cannot form stable adsorbed layer, and then can not obtain stable detection signal.If higher than 60min, although still cleaning performance can be ensured, do not meet the on-line continuous test request of instrument, so the preferred 5-60min of scavenging period of the present invention.If the concentration OD600 of microorganism is lower than 0.01 in cleaning fluid, then the organic content in cleaning fluid is too low, cannot form the organic adsorption layer that electrode can be suppressed to pollute further, and then cannot obtain and detect data reliably.If higher than 0.2, then in During Process of Long-term Operation, easily accumulate too much organism at electrode surface and form microbial film, affecting the output performance steady in a long-term of microelectrode detecting device.
In the present invention, the kind of the microorganism of adding does not limit, it can be single culture, also can be the mixing of multiple bacterial classification, general microorganism is obtained by water sample Culture in situ to be measured, Culture in situ method is prior art, and in cleaning fluid, the kind of microorganism can be the same or different with the kind of the microorganism be added in water sample to be measured, preferably identical; Electron mediator is the derivant of the potassium ferricyanide, dimethyl diaminophenazine chloride, ferrocene or ferrocene; The material of microelectrode is the noble metals such as gold, platinum, is not particularly limited.
The present invention is further illustrated below in conjunction with embodiment.
Embodiment 1
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 0.2, and the concentration of the potassium ferricyanide is 2g/L;
Step 3, get partial test sample, dilute with water sample to be measured, be 0.01 to microorganism concn OD600, obtain cleaning fluid;
Step 4, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain control sample, in control sample, microorganism concn OD600 is 0.2, and the concentration of the potassium ferricyanide is 2g/L;
Step 5, with cleaning fluid cleaning pick-up unit microelectrode (platinum microelectrodes of 20 μm) 60min, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 6, run pick-up unit continuously, repeat step 5, to detecting 400 times altogether.
The water sample to be measured that embodiment 1 adopts is deionized water, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).Fig. 1 is the ratio that embodiment 1 difference detects the limiting current of test specimens under number of times and control sample, as can be seen from the figure, the deviation of the detection electric current after continuous 350 tests is less than 2%, illustrate that present invention reduces test specimens in testing process, to the pollution of working electrode, obtains stable current signal and export.
Embodiment 2
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 3, get partial test sample, dilute with water sample to be measured, be 0.01 to microorganism concn OD600, obtain cleaning fluid;
Step 4, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain control sample, in control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 5, with cleaning fluid cleaning pick-up unit microelectrode (the gold dish microelectrodes of 25 μm) 5min, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 6, run pick-up unit continuously, repeat step 5, to detecting 100 times altogether.
The water sample to be measured that embodiment 2 adopts is deionized water, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).Fig. 2 is real-time current value and the relation curve of detection time of embodiment 2 record, Fig. 3 is the partial enlarged drawing of Fig. 2, be the control sample after cleaning fluid, cultivation, the test specimens after cultivation once continues to flow through the current signal that microelectrode detecting device produces successively, as can be seen from Fig. 2-3, pollute although test specimens can produce electrode when flowing through microelectrode, but after repeatedly testing, the current signal difference of control sample and test specimens is still less than 3%.
Embodiment 3
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 1, and the concentration of the potassium ferricyanide is 3g/L;
Step 3, get partial test sample, dilute with water sample to be measured, be 0.2 to microorganism concn OD600, obtain cleaning fluid;
Step 4, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain control sample, in control sample, microorganism concn OD600 is 1, and the concentration of the potassium ferricyanide is 3g/L;
Step 5, with cleaning fluid cleaning pick-up unit microelectrode (the gold dish microelectrodes of 25 μm) 60min, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 6, run pick-up unit continuously, repeat step 5, to detecting 120 times altogether.
The water sample to be measured that embodiment 3 adopts is Songhuajiang River Water, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).After tested, the deviation of the detection electric current after continuous 120 tests is less than 2%.
Embodiment 4
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 6, and the concentration of the potassium ferricyanide is 15g/L;
Step 3, get partial test sample, dilute with water sample to be measured, be 0.1 to microorganism concn OD600, obtain cleaning fluid;
Step 4, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain control sample, in control sample, microorganism concn OD600 is 6, and the concentration of the potassium ferricyanide is 15g/L;
Step 5, with cleaning fluid cleaning pick-up unit microelectrode (the gold dish microelectrodes of 25 μm) 60min, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 6, run pick-up unit continuously, repeat step 5, to detecting 100 times altogether.
The water sample to be measured that embodiment 4 adopts is her logical river, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).After tested, the deviation of the detection electric current after continuous 100 tests is less than 2%.
Comparative example 1
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 3, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain control sample, in control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 4, be the potassium ferricyanide of 2g/L with concentration be cleaning fluid, first with microelectrode (platinum microelectrodes of the 20 μm) 5min of cleaning fluid cleaning pick-up unit, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 5, run pick-up unit continuously, repeat step 4, to detecting 100 times altogether.
The water sample to be measured that comparative example 1 adopts is deionized water, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).Fig. 4 is cleaning fluid in comparative example 1, cultivate after control sample, cultivate after test specimens once continue to flow through microelectrode successively, the real-time current value of detection device records.As can be seen from the figure, the deviation of control sample and test specimens is more than 10%.
Comparative example 2
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 3, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain control sample, in control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 4, with alkaline solution (0.1M KOH containing 0.5% song draw logical) for cleaning fluid, first with microelectrode (platinum microelectrodes of the 20 μm) 5min of cleaning fluid cleaning pick-up unit, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 5, run pick-up unit continuously, repeat step 4, to detecting 100 times altogether.
The water sample to be measured that comparative example 2 adopts is deionized water, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).After tested, the deviation of control sample and test specimens is more than 10%.
Comparative example 3
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 3, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain control sample, in control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 4, with acid solution (0.1M KOH containing 0.5% song draw logical) for cleaning fluid, first with microelectrode (platinum microelectrodes of the 20 μm) 5min of cleaning fluid cleaning pick-up unit, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 5, run pick-up unit continuously, repeat step 4, to detecting 100 times altogether.
The water sample to be measured that comparative example 3 adopts is deionized water, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).After tested, the deviation of control sample and test specimens is more than 30%.
Comparative example 4
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 3, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain control sample, in control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 4, with the deionized water of 60 DEG C for cleaning fluid, first with microelectrode (platinum microelectrodes of the 20 μm) 5min of cleaning fluid cleaning pick-up unit, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 5, run pick-up unit continuously, repeat step 4, to detecting 100 times altogether.
The water sample to be measured that comparative example 4 adopts is deionized water, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).Fig. 5 is cleaning fluid in comparative example 4, cultivate after control sample, cultivate after test specimens once continue to flow through microelectrode successively, the real-time current value of detection device records.As can be seen from the figure, the deviation of control sample and test specimens is more than 10%.
Comparative example 5
Step one, get water sample to be measured and carry out Culture in situ, obtain microorganism;
Step 2, get water sample to be measured again, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain test specimens, in test specimens, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 3, remove ionized water, add the microorganism that the potassium ferricyanide and step one are cultivated wherein, obtain in control sample, in control sample, microorganism concn OD600 is 0.1, and the concentration of the potassium ferricyanide is 2g/L;
Step 4, with the deionized water of 60 DEG C for cleaning fluid, first use cleaning fluid with microelectrode (platinum microelectrodes of the 20 μm) 5min of the form of high voltage pulse (being greater than 2kg) cleaning pick-up unit, then the control sample after microbial cultivation device is cultivated and test specimens is detected successively with pick-up unit, the real-time current value of micro-electric current in record testing process;
Step 5, run pick-up unit continuously, repeat step 4, to detecting 100 times altogether.
The water sample to be measured that comparative example 5 adopts is deionized water, and pick-up unit is the disclosed pick-up unit detecting water body toxicity of Chinese patent (publication number 102520047A).Fig. 6 is cleaning fluid in comparative example 5, cultivate after control sample, cultivate after test specimens once continue to flow through microelectrode successively, the real-time current value of detection device records.As can be seen from the figure, the deviation of control sample and test specimens is more than 10%.
It should be noted that, for meeting single contrast principle, the water sample to be measured that comparative example 1-5 and embodiment 2 adopt is identical.The testing result of comparative example 2 and comparative example 1-5 can be found out, after adopting cleaning method cleaning electrode of the present invention, reduce test specimens in testing process, to the pollution of working electrode, stable current signal can be obtained and export, even if repetitive measurement deviation is also very little.
Obviously, the explanation of above embodiment just understands method of the present invention and core concept thereof for helping.It should be pointed out that the those of ordinary skill for described technical field, under the premise without departing from the principles of the invention, can also carry out some improvement and modification to the present invention, these improve and modify and also fall in the protection domain of the claims in the present invention.
Claims (6)
1. the cleaning method of the microelectrode of water body toxicity pick-up unit, is characterized in that, with cleaning fluid cleaning more than electrode 5min;
Described cleaning fluid is the mixed solution of water sample to be measured and microorganism, and in cleaning fluid, the concentration OD600 of microorganism is 0.02-0.1.
2. the cleaning method of the microelectrode of water body toxicity pick-up unit according to claim 1, is characterized in that, described cleaning fluid adds the water sample to be measured after microorganism by dilution and obtains, and described diluting solvent is water sample to be measured.
3. the cleaning method of the microelectrode of water body toxicity pick-up unit according to claim 1, is characterized in that, also containing electron mediator in described cleaning fluid.
4. the cleaning method of the microelectrode of water body toxicity pick-up unit according to claim 3, is characterized in that, described cleaning fluid adds the water sample to be measured after electron mediator and microorganism by dilution and obtains, and described diluting solvent is water sample to be measured.
5. the cleaning method of the microelectrode of water body toxicity pick-up unit according to claim 3, is characterized in that, described electron mediator is the derivant of the potassium ferricyanide, dimethyl diaminophenazine chloride, ferrocene or ferrocene.
6., according to the cleaning method of the microelectrode of the water body toxicity pick-up unit of claim 1-5 described in any one, it is characterized in that, described scavenging period is 5-60min.
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