CN108344618B - Quick malachite green detection method - Google Patents
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- CN108344618B CN108344618B CN201810020745.2A CN201810020745A CN108344618B CN 108344618 B CN108344618 B CN 108344618B CN 201810020745 A CN201810020745 A CN 201810020745A CN 108344618 B CN108344618 B CN 108344618B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
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- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
Abstract
The invention discloses a method for rapidly detecting malachite green, which comprises the steps of taking sodium sulfite or sodium thiosulfate as a sensitizer, adding a malachite green solution into a buffer solution containing the sensitizer, standing an electrode in the sensitizer buffer solution containing the malachite green for open-circuit enrichment, carrying out addition reaction on the malachite green and the sodium sulfite to form an intermediate product, enriching the intermediate product onto the electrode, detecting by using a square wave anodic stripping voltammetry to obtain a standard curve of stripping current changing along with the concentration of the malachite green solution, and judging the concentration of the malachite green solution to be detected, namely the content of the malachite green by using the square wave anodic stripping voltammetry and the standard curve. The preparation and detection processes of the detection method reagent do not need expensive instruments and reagents, the cost is low, the operation is simple, the detection sensitivity is high, the test result is stable, and the market application prospect is good.
Description
Technical Field
The invention relates to the field of organic compound detection and analysis, in particular to a rapid malachite green detection method.
Background
Malachite green is a triphenylmethane type green dye, commonly known as alkaline green, aniline green and the like, and is widely used in leather industry, paper making industry, textile printing and dyeing industry. Meanwhile, malachite green is also an excellent sterilization and insecticide, and has good killing effect on various common aquatic diseases such as saprolegniasis, branchiomycosis, ichthyophthiriasis and the like of fish, shrimps and the like; however, malachite green is rapidly metabolized into colorless malachite green in fish bodies, and the colorless malachite green has higher toxicity, is easy to remain, has teratogenic and carcinogenic effects, and is forbidden for aquaculture prohibited by most countries. However, malachite green is low in price, excellent in effect and no corresponding substitute product exists at present, so that the malachite green is forbidden frequently. The conventional detection method for malachite green mainly focuses on two types, one type is a physicochemical detection method, the other type is an enzyme-linked immunoassay method, the methods are qualitative detection methods, or large instruments and professional operators are required, or false positive problems exist, and the price is high, so that a simple, effective, high-specificity and low-cost detection method needs to be invented.
Disclosure of Invention
In order to solve the problems, the invention provides a method for rapidly detecting malachite green, which comprises the steps of taking sodium sulfite or sodium thiosulfate as a sensitizer, adding a malachite green solution into a buffer solution containing the sensitizer, standing an electrode in the sensitizer buffer solution containing the malachite green for open circuit enrichment, carrying out addition reaction on the malachite green and the sodium sulfite to form an intermediate product, enriching the intermediate product onto the electrode, detecting by using a square wave anodic stripping voltammetry to obtain a standard curve of stripping current changing along with the concentration of the malachite green solution, and judging the concentration of the malachite green solution to be detected, namely the content of the malachite green by using the square wave anodic stripping voltammetry and the standard curve. The preparation and detection processes of the detection method reagent do not need expensive instruments and reagents, the cost is low, the operation is simple, the detection sensitivity is high, the test result is stable, and the market application prospect is good.
The technical scheme of the invention is as follows:
a rapid malachite green detection method is characterized by comprising the following steps:
1) adding a sodium sulfite solution or a sodium thiosulfate solution into an acetate buffer solution or a phosphate buffer solution, and uniformly mixing to obtain a sensitizer mixed solution;
2) immersing an electrode connected to an electrochemical workstation into the sensitizer mixed solution in the step 1), and selecting an anodic stripping voltammetry on the electrochemical workstation to perform electrode activation on the electrode;
3) after the electrode activation is finished, adding a malachite green solution into the sensitizer mixed solution soaked with the electrode in the step 2), and standing for 5-10 minutes;
4) after standing, detecting by using an anodic stripping voltammetry method on an electrochemical workstation to obtain a stripping voltammogram of the malachite green in the malachite green solution in the step 3);
5) repeating the steps 1) to 4), measuring dissolution voltammograms of malachite green in malachite green solutions with different concentrations, and drawing a standard curve of dissolution current along with the change of the concentration of the malachite green solution according to the voltammograms;
6) and (3) repeating the steps 1) to 4) to determine a dissolution voltammogram of the malachite green in the malachite green solution to be detected, obtaining dissolution current according to the voltammogram, and obtaining the concentration of the malachite green solution to be detected by utilizing the standard curve of the step 5).
The acetate buffer solution is an acetic acid solution of acetate, and the pH value is 4.0-5.0.
In the step 1), the molar ratio of sodium sulfite or sodium thiosulfate to acetate in an acetate buffer solution is 0.002: 1.
Step 1) adding 2 mu L of sodium sulfite solution or sodium thiosulfate solution with the concentration of 0.1mol/L into 1mL of acetate buffer solution with the concentration of 0.1mol/L, and uniformly mixing to obtain the sensitizer mixed solution.
The electrodes are screen-printed electrodes.
The screen printing electrode is a three-electrode screen printing electrode plate.
The anodic stripping voltammetry is square wave anodic stripping voltammetry.
The concentrations of the malachite green solution in the step 5) and the step 6) are 36ppb, 72ppb, 108ppb, 144ppb, 180ppb, 225ppb and 270ppb in sequence.
During the electrode activation process, the detection potential is 0.3V-0.7V, the voltage increment is 4mV, the amplitude is 0.025V, and the frequency is 15 Hz.
When the anodic stripping voltammetry on the electrochemical workstation is selected for detection in the step 4), the detection potential is 0.3V-0.7V, the voltage increment is 4mV, the amplitude is 0.025V, and the frequency is 15 Hz.
The invention has the technical effects that:
according to the rapid detection method of malachite green, sodium sulfite or sodium thiosulfate and malachite green are subjected to addition reaction in an acetate buffer solution environment to form an intermediate, and the specific reaction is as follows:
the reaction intermediate is as follows:
the reaction intermediate can reduce the detection potential of malachite green, the electrode is placed in the sensitization mixed liquid in which the malachite green is dissolved, the generated reaction intermediate is easy to enrich to the surface of the electrode, and therefore the effect of signal amplification is achieved, particularly, the electrode enrichment process of the intermediate is open circuit enrichment, namely, the electrode is subjected to standing enrichment under the condition that a power supply is not switched on, and the process improves the detection sensitivity to a certain extent. After the electrode is stood, certain reaction intermediates are enriched, and then the anode stripping voltammetry detection is carried out by using a portable electrochemical workstation, so that a detection platform capable of detecting the malachite green on site simply, quickly and highly sensitively is constructed.
In addition, in the detection process, a screen printing electrode is adopted, so that the electrode is low in cost, easy and convenient to operate and very stable in test performance, and can be thrown when used; the square wave anodic stripping voltammetry is adopted for detection, the voltammetry curve is clear, and the change trend is obvious.
The detection method has the advantages that the malachite green detection limit can be as low as 100nM, the detection time is short, expensive reagents such as antibodies and the like required in the construction of a sensor in the conventional detection method are not required, only trace sulfite or thiosulfate is required, and the detection cost is greatly reduced; the method is simple to operate and high in detection sensitivity; meanwhile, the construction and use processes of the sensor do not need to depend on large-scale instruments and professional operators, and the sensor can be widely applied to field quantitative detection in rural areas with deficient resources and developing countries.
Drawings
FIG. 1 is a voltammogram showing the dissolution of malachite green in a malachite green solution having a concentration of 36ppb to 270ppb in examples of the present invention.
FIG. 2 is a graph of the linear fit of the concentration of 36ppb-270ppb malachite green solution to the dissolution current in the example of the invention.
Detailed Description
The technical solution of the present invention is described in detail by the following specific examples, but the scope of the present invention is not limited by these examples.
The electrochemical workstation used in the embodiment of the present invention is a portable electrochemical workstation.
The ppm of the invention is mg/L, and the ppb is mug/L.
Example 1
The embodiment provides a malachite green rapid detection method, which comprises the following steps:
1) adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium thiosulfate solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a 36ppb malachite green solution into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz to obtain the voltammetry curve.
Example 2
The embodiment provides a malachite green rapid detection method, which comprises the following steps:
1) adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium sulfite solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a 36ppb malachite green solution into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz to obtain a voltammetry curve, which is shown as a curve in figure 1.
By analyzing examples 1 and 2, and selecting example 2 as a basis, various concentrations of malachite green solutions were determined.
Example 3
The embodiment provides a malachite green rapid detection method, which comprises the following steps:
1) adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium sulfite solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a 72ppb malachite green solution into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz to obtain a voltammetry curve, as shown by a curve b in figure 1.
Example 4
The embodiment provides a malachite green rapid detection method, which comprises the following steps:
1) adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium sulfite solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a 108ppb malachite green solution into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz to obtain a voltammetry curve, which is shown as a curve c in figure 1.
Example 5
The embodiment provides a malachite green rapid detection method, which comprises the following steps:
1) adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium sulfite solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a 144ppb malachite green solution into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz to obtain a voltammetry curve, as shown by a curve d in figure 1.
Example 6
The embodiment provides a malachite green rapid detection method, which comprises the following steps:
1) adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium sulfite solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a solution of malachite green with the concentration of 180ppb into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating the malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz to obtain a voltammetry curve, as shown by the curve e in figure 1.
Example 7
The embodiment provides a malachite green rapid detection method, which comprises the following steps:
1) adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium sulfite solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a 225ppb malachite green solution into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: the voltammetry curve is obtained by detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz, and is shown as the f curve in figure 1.
Example 8
The embodiment provides a malachite green rapid detection method, which comprises the following steps:
1) adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium sulfite solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a 270ppb malachite green solution into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz to obtain a voltammetry curve, which is shown as a g curve in figure 1.
According to examples 2 to 8, measured concentrations were 36ppb, 72ppb, 108ppb, 144ppb, 180ppb, 225ppb, and 270ppb, respectively, of the dissolution voltammogram of malachite green in the solution of malachite green, and a standard curve of the dissolution current as a function of the concentration of the malachite green solution, i.e., a linear fit graph of the concentration of the malachite green solution and the dissolution current, was plotted on the basis of the voltammogram, as shown in fig. 2.
Example 9
1) Adding 1mL of sodium acetate buffer solution with the concentration of 0.1mol/L and the pH value of 4.5 into 2 mu L of 0.1mol/L sodium sulfite solution, and uniformly oscillating to obtain sensitizer mixed solution;
2) connecting a screen printing electrode with a portable electrochemical workstation, immersing the screen printing electrode into the sensitizer mixed solution prepared in the step 1), selecting a square wave anodic stripping voltammetry, activating a potential window of 0.3V-0.7V, increasing the voltage by 4mV, increasing the amplitude by 0.025V and performing the activation of the screen printing electrode at the frequency of 15 Hz;
3) after activation, adding a malachite green solution to be detected into the sensitizer mixed solution soaked with the screen printing electrode in the step 2), pretreating the malachite green, and standing for 5 minutes;
4) after standing, forming an intermediate by malachite green and sodium sulfite, and detecting a solution system by adopting a square wave anodic stripping voltammetry, wherein the parameters are set as follows: detecting the potential of 0.3V-0.7V, the voltage increment of 4mV, the amplitude of 0.025V and the frequency of 15Hz to obtain a voltammetry curve, obtaining the dissolution current of 0.2 muA under the concentration according to the voltammetry curve, and obtaining the concentration of the malachite green to be detected to be 201ppb according to the dissolution current of 0.2 muA and corresponding to the standard curve shown in figure 2.
The detection method disclosed by the invention does not need expensive reagents such as antibodies and the like required in the traditional detection method when a sensor is constructed, only needs trace sulfite or thiosulfate, and greatly reduces the detection cost; the detection time is short, the operation is simple, and the detection sensitivity is high; the screen printing electrode has low cost, is thrown when being used, is simple and convenient to operate and has very stable test performance; the square wave anodic stripping voltammetry is adopted for detection, and the characteristics of the voltammetry curve are obvious; meanwhile, the construction and use processes of the detection method sensor do not need to depend on large-scale instruments and professional operators, and the method can be widely applied to field quantitative detection in resource-deficient rural areas and developing countries.
It should be noted that the above examples are only illustrative and not restrictive, and that the present invention can be modified in any way without departing from the spirit of the present invention, and the technical scope of the present invention is to be determined from the changes of sulfite concentration, sodium hydroxide concentration, activation potential, activation time, pretreatment potential, pretreatment time and detection parameters.
Claims (10)
1. A rapid malachite green detection method is characterized by comprising the following steps:
1) adding a sodium sulfite solution or a sodium thiosulfate solution into an acetate buffer solution, and uniformly mixing to obtain a sensitizer mixed solution;
2) immersing an electrode connected to an electrochemical workstation into the sensitizer mixed solution in the step 1), and selecting an anodic stripping voltammetry on the electrochemical workstation to perform electrode activation on the electrode;
3) after the electrode activation is finished, adding a malachite green solution into the sensitizer mixed solution soaked with the electrode in the step 2), and standing for 5-10 minutes;
4) after standing, detecting by using an anodic stripping voltammetry method on an electrochemical workstation to obtain a stripping voltammogram of the malachite green in the malachite green solution in the step 3);
5) repeating the steps 1) to 4), measuring dissolution voltammograms of malachite green in malachite green solutions with different concentrations, and drawing a standard curve of dissolution current along with the change of the concentration of the malachite green solution according to the voltammograms;
6) and (3) repeating the steps 1) to 4) to determine a dissolution voltammogram of the malachite green in the malachite green solution to be detected, obtaining dissolution current according to the voltammogram, and obtaining the concentration of the malachite green solution to be detected by utilizing the standard curve of the step 5).
2. The method for rapidly detecting malachite green as claimed in claim 1, wherein the acetate buffer solution is an acetic acid solution of acetate, and the pH value is 4.0-5.0.
3. The method for rapidly detecting malachite green as claimed in claim 2, wherein in step 1), the molar ratio of sodium sulfite or sodium thiosulfate to acetate in acetate buffer solution is 0.002: 1.
4. The method for rapidly detecting malachite green according to claim 3, characterized in that in step 1), 2 μ L of sodium sulfite solution or sodium thiosulfate solution with concentration of 0.1mol/L is added into 1mL of acetate buffer solution with concentration of 0.1mol/L, and the mixture is mixed uniformly to obtain the sensitizer mixture.
5. The method for rapidly detecting malachite green as claimed in claim 1, wherein the electrodes are screen-printed electrodes.
6. The method for rapidly detecting malachite green according to claim 5, characterized in that the screen-printed electrode is a three-electrode screen-printed electrode.
7. The method for rapidly detecting malachite green according to claim 1, characterized in that the anodic stripping voltammetry is square wave anodic stripping voltammetry.
8. The method for rapidly detecting malachite green as claimed in claim 1, characterized in that the concentration of malachite green solution in step 5) is 36ppb, 72ppb, 108ppb, 144ppb, 180ppb, 225ppb, 270ppb in sequence.
9. The method for rapidly detecting malachite green according to claim 1, characterized in that during the activation of the electrodes, the detection potential is 0.3V-0.7V, the voltage increment is 4mV, the amplitude is 0.025V, and the frequency is 15 Hz.
10. The method for rapidly detecting malachite green according to claim 1, characterized in that, when anodic stripping voltammetry on an electrochemical workstation is selected in the step 4) for detection, the detection potential is 0.3V-0.7V, the voltage increment is 4mV, the amplitude is 0.025V, and the frequency is 15 Hz.
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| CN101949846A (en) * | 2010-08-11 | 2011-01-19 | 昆明理工大学 | Leucomalachite green detection method |
| CN102004099A (en) * | 2010-09-17 | 2011-04-06 | 大连理工大学 | Method for detecting malachite green through electrochemiluminescence |
| CN106556593A (en) * | 2016-11-18 | 2017-04-05 | 常州大学 | A kind of method of fixing tripyridyl ruthenium and detecting malachite green through electrochemiluminescence |
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