CN108459062B - Nano copper oxide modified electrode and method for analyzing tilmicosin by using modified electrode - Google Patents
Nano copper oxide modified electrode and method for analyzing tilmicosin by using modified electrode Download PDFInfo
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Abstract
The invention discloses a method for modifying an electrode by nano copper oxide, which comprises the steps of preparing nano copper oxide, preparing nano copper oxide suspension, modifying a glassy carbon electrode and the like. The invention also discloses a method for analyzing timixacin by using the modified electrode, which comprises the following steps: preparing a tilmicin mother solution; preparing a buffer solution; providing the nano copper oxide modified electrode; adding the tilmicin solution into a buffer solution, using the nano copper oxide modified electrode in the step as a working electrode, using a platinum electrode as an auxiliary electrode and using a calomel electrode as a corresponding reference electrode to jointly form a three-electrode system, and detecting the content of tilmicin by using a cyclic voltammetry. According to the invention, the nano copper oxide modified electrode is obtained by a method of modifying a glassy carbon electrode by nano copper oxide, and the modified electrode is used for analyzing the tilifloxacin, so that the analysis result has good reproducibility and high stability.
Description
Technical Field
The invention relates to the field of nano materials, in particular to a method for modifying an electrode by using nano copper oxide and application of the modified electrode in tilmisartan analysis.
Background
The tilmicin is a valuable antibiotic drug developed and produced in 90 years of the 20 th century. The drug is a product with higher activity obtained by amination of aldehyde group after desugarization of tylosin, and the reaction takes formic acid as a catalyst, so that the scheme has high yield. At present, the main detection methods for timixacin are classified into high performance liquid chromatography, ultraviolet spectrophotometry, voltammetry and nuclear magnetic resonance. These methods do not fully satisfy the requirements in terms of reproducibility, stability, cost, etc., and thus a novel analytical method is required for the analytical detection of temifloxacin.
Disclosure of Invention
The invention provides a method for modifying an electrode by using nano copper oxide with good reproducibility and high stability and a method for analyzing timixacin by using the modified electrode.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a method for modifying an electrode by nano copper oxide comprises the following steps:
s10, preparing nano copper oxide: adding polyethylene glycol into the copper acetate solution to form a solution A; adding polyethylene glycol into a sodium carbonate solution, dissolving to form a solution B, uniformly dripping the solution B into the solution A under the ultrasonic condition, performing ultrasonic treatment for a proper time, performing magnetic stirring for 30 minutes at 78 ℃ to obtain a black precipitate, centrifuging the black precipitate, repeatedly washing the black precipitate for multiple times, and performing vacuum drying to obtain a copper oxide nano material;
s20, preparing a nano copper oxide suspension: weighing copper oxide nanopowder, placing the copper oxide nanopowder in a sample tube, dripping distilled water into the sample tube to enable the mass ratio of the copper oxide nanopowder to the water to be 1:200, stirring for 5-10 minutes, then adding 1.00mL of 1% PVP into the suspension, and finally placing the sample tube in an ultrasonic cleaner to vibrate and disperse for 30min for later use; shaking for 15-20min before each use so that the nano copper oxide particles can be uniformly dispersed;
s30, modifying glassy carbon electrode: firstly, polishing and grinding a glassy carbon electrode on aluminum oxide powder, then placing the glassy carbon electrode in a beaker filled with dilute sulfuric acid, shaking and cleaning for 5-10min, then taking out and cleaning with distilled water, and finally placing at room temperature for air drying; and (3) sucking 10 microliters of the uniformly dispersed copper oxide nano turbid liquid by using a micro syringe, dripping and coating the surface of an electrode core of the glassy carbon electrode by using a dripping method, and then drying the electrode core under infrared light to obtain the modified electrode of which the surface is filled with a thin layer of nano copper oxide particles.
A method for analyzing timixacin by using a modified electrode, comprising the following steps:
s40, preparing a tilmicafloxacin mother solution;
s50, preparing a buffer solution;
s60, providing the nano copper oxide modified electrode;
s70, adding the tilmicin solution into a buffer solution, using the nano copper oxide modified electrode in the step S60 as a working electrode, a platinum electrode as an auxiliary electrode and a calomel electrode as corresponding reference electrodes to jointly form a three-electrode system, and detecting the tilmicin content by using a cyclic voltammetry.
Wherein in step S50, the buffer is phosphate buffer.
In step S50, the pH of the phosphate buffer is 7.52.
In step S70, the scanning rate of the nano-copper oxide modified glassy carbon electrode is 0.10V/S.
Wherein the step S40 specifically includes the following steps:
accurately weighing a certain mass of tilmicafloxacin technical product, placing the tilmicafloxacin technical product in a beaker, adding water for dissolving, and then fixing the volume to a volumetric flask to form 0.2 × 10-2And (3) placing the tilmicin mother liquor in a dark place for later use.
Wherein the step S50 specifically includes the following steps:
s51, preparing (1)0.4mol/L NaH2PO4: weighing NaH2PO4·2H2Dissolving 31.2g of O in 500.00ml of distilled water;
s52, preparing (2)0.4mol/L Na2HPO4: weighing Na2HPO4·12H2Dissolving O71.6 g in 500.00ml of distilled water;
s53, mixing the above (1) and (2) according to different ratios to obtain phosphate buffer solution with required pH value, and measuring by a pH instrument to prepare required mixed buffer solution.
According to the invention, the nano copper oxide modified electrode is obtained by a method of modifying a glassy carbon electrode by nano copper oxide, and the modified electrode is used for analyzing the tilifloxacin, so that the analysis result has good reproducibility and high stability.
Drawings
FIG. 1 is a CV curve diagram of timifloxacin on a bare glassy carbon electrode and a nano copper oxide modified glassy carbon electrode in an embodiment of the invention.
FIG. 2 is a comparison graph of electrochemical response behaviors of timixacin in different buffers in the example of the present invention.
FIG. 3 is a graph of reduction current versus pH for the examples of the present invention.
FIG. 4 is a graph illustrating the relationship between the reduction current and the scan rate according to an embodiment of the present invention.
FIG. 5 is a DPV graph of different concentrations of temifloxacin under optimal conditions in the examples of the present invention.
FIG. 6 is a graph of reduction current versus concentration of tilmifloxacin in an example of the present invention.
Detailed Description
The invention is further described below with reference to the accompanying drawings and examples.
In this embodiment, the method for modifying the electrode with the nano copper oxide includes three steps of preparing the nano copper oxide, preparing a nano copper oxide suspension, and modifying the glassy carbon electrode.
The method for preparing the nano copper oxide comprises the following steps: adding PEG (polyethylene glycol) into the copper acetate solution to form a solution A; and adding PEG (polyethylene glycol) into the sodium carbonate solution, and dissolving to form a solution B. And then uniformly dripping the solution B into the solution A under the ultrasonic condition, carrying out ultrasonic treatment for a proper time, finally carrying out magnetic stirring for 30 minutes at 78 ℃ to obtain a black precipitate, centrifuging the black precipitate, repeatedly washing the black precipitate for multiple times, and carrying out vacuum drying to obtain the copper oxide nano material with the particle size of about 50-70 nm. FIG. 1 shows the scanning result of the electron lens of the nano-copper oxide product of this embodiment.
Preparing a nano copper oxide suspension: accurately weighing 0.0100g of copper oxide nanopowder, placing in a sample tube, adding 2.00mL of distilled water dropwise, slightly stirring for 5-10min, adding 1.00mL of 1% PVP (polyvinylpyrrolidone) into the suspension, and placing the sample tube in an ultrasonic cleaner for shaking and dispersing for 30min for later use. Shaking for 15-20min before each use so that the nano copper oxide particles can be uniformly dispersed.
The modification method of the glassy carbon electrode comprises the following steps: firstly, polishing and grinding a glassy carbon electrode on aluminum oxide powder, then placing the glassy carbon electrode in a beaker filled with dilute sulfuric acid, shaking and cleaning for 5-10min, then taking out and cleaning with distilled water, and finally placing at room temperature for air drying. And sucking 10.00 microliters of the uniformly dispersed copper oxide nano turbid liquid by using a micro syringe, dripping and coating the surface of an electrode core of the glassy carbon electrode by adopting a dripping method, and then drying the electrode core under infrared light to obtain the modified electrode of which the surface is filled with a thin layer of nano copper oxide particles.
In this example, the analysis of timixacin with the modified electrode includes the following steps.
S40, preparing the tilmicafloxacin mother liquor
Accurately weighing 0.1738g of tilmicin original drug, placing the original drug in a beaker, adding water to dissolve the original drug, and then fixing the volume to a 100.00mL volumetric flask to form 0.2 × 10-2And (3) mol/L of the tilmicin mother liquor. And placing in the dark for standby.
S50, preparing a buffer solution
In order to study the influence of different buffer base solutions on the electrochemical response of tilifloxacin, phosphate buffer solution, BR buffer solution, sodium tetraborate buffer solution and NH were selected in this example3-NH4Cl buffer and Na2CO3-NaHCO3Five buffers such as buffer.
The principle of the embodiment is as follows: a glassy carbon electrode modified by nano copper oxide is taken as a working electrode, a platinum electrode is taken as an auxiliary (counter) electrode, and a 222 type calomel electrode is taken as a corresponding reference electrode, so that a three-electrode system is formed together. The response of various drugs is qualitatively searched under various buffer base solutions by Cyclic Voltammetry (CV), then the optimal buffer solution, the optimal pH value and the scanning rate are determined, and after all the responses are determined, a standard curve, the reproducibility, the lowest detection limit and the standard addition recovery condition are quantitatively made through DPV, so that the corresponding electrochemical behavior of the drugs is presumed. The experimental data were recorded and saved with an electrochemical workstation.
Comparison of bare electrode with modified electrode
In fig. 1, a is the response peak current of the modified electrode to tilmisartan; b is the response peak current of the bare glassy carbon electrode to tilmicosin; and C is the response curve of the modified electrode to phosphate buffer. From fig. 1 it can be concluded that phosphate is not interfering in the response interval, which is due to the electrochemical behavior of temifloxacin. The comparison of the curves A and B shows that the copper oxide modified electrode has stronger response than a bare electrode, which shows that the nano modifier copper oxide plays a role in catalyzing the electrochemical behavior of the tilifloxacin.
Selection of buffer
FIG. 2 shows phosphate buffer, BR buffer, sodium tetraborate buffer, NH3-NH4Cl buffer and Na2CO3-NaHCO3Effects of five buffers, e.g., buffer, on electrochemical response of timifloxacin. As can be seen from FIG. 2, the response effect of the phosphate buffer (curve A) is more prominent, and the peak effect is relatively better, so that the phosphate buffer is determined to be the optimal buffer for electrochemical response of tilmicin. .
Determination of the optimum pH
Aiming at the selection of pH, five groups of parallel experiments are carried out, phosphate buffer solutions with constant concentration and pH of 6.48, 7.03, 7.52, 8.03 and 8.34 are prepared, the oxidation potential of phosphate buffer solutions with different pH values under the nano copper oxide modified glassy carbon electrode is measured, and graph 3 is drawn according to the relationship between the pH value and the oxidation potential corresponding to the current magnitude. From the relationship between PH and oxidation current, the optimum PH of the phosphoric acid base solution was 7.52.
Determination of optimal scan rate
Preparing a tilmisartan standard solution taking phosphate base solution with pH of 7.52 as a buffer solution, detecting under a three-electrode system taking a copper oxide nano modified electrode as a working electrode, and taking scanning rates of 0.06, 0.08, 0.10, 0.12 and 0.14V/s to respectively perform multiple times of measurement. From FIG. 4, it can be seen that when the scan rate is equal to 0.10V/s, the oxidation peak current is strongest, and the response effect is most obvious, and in summary, the optimal scan rate is 0.10V/s.
Linear range and detection limit of temifloxacin
Under the best experimental conditions (pH 7.52, scan rate 0.10V/s), DPV was used for different concentrations (FIG. 5: A is 1.2 × 10)-7mol/L, B is 2.4 × 10-7mol/L, C is 3.6 × 10-7mol/L, D is 4.8 × 10-7mol/L) and has a concentration gradientThe tilmicosin standard solution of degree was analyzed by detection, a linear curve of concentration and oxidation current was prepared according to FIG. 6, and it was found from FIG. 6 that the concentration was 1.2 × 10-7mol/L~4.8×10-7The mol/L ranges all have linear relations, the linear equation is shown as y which is 70.16x + E-06, the correlation linear coefficient is 0.9826, according to the detection limit formula LOD which is 3 sigma/S (sigma represents blank standard deviation, and S represents the slope of the correction curve), the detection limit can be calculated to be 7.9 × 10-8mol/L。
Reproducibility and stability of nano modified electrode
Stability: the modified electrode is placed at room temperature for a week, and then the detection of substances with the same content is carried out, the result is reduced by 0.85% in the same ratio, which indicates that the stability of the modified electrode is good.
Reproducibility the concentration of the modified electrode was determined to be 2.5 × 10 in 10 repetitions-8From the results of 10 times of measurement, the relative deviation of peak current of the tilifloxacin solution of mol/L is maintained to be about 4.7% -6.9%, and the experimental result shows that the reproducibility of the modified electrode is relatively good.
Interference experiment of nano modified electrode
The influence of auxiliary materials of the tilmicafloxacin on the electrochemical response is investigated, and the result shows that the dry starch, the lubricant, the polyethylene glycol and the like which are more than 10 times do not interfere with the experimental result. Because other antibiotic drugs such as cephalosporins and penicillins are often added into the temifloxacin, a large number of experiments exclude that the modified electrode responds to the drugs; and the interference degree of antioxidant substances, organic solvents and the like contained in the medicament is also slightly influenced, and the result shows that the anti-interference performance of the copper oxide nano modified electrode is very strong.
Timicacein content determination and standard recovery rate calculation
And (3) the recovery amount of the timifloxacin gazette: taking three tilmicin sample solutions with unchanged concentrations, adding 5mL of phosphate buffer solution, and carrying out analysis and detection under the optimal condition; then, a certain amount of tilmicosin standard solution with the same volume is added, and then the analysis and detection are carried out under the optimal condition. And finally, subtracting the calculated value before the labeling from the calculated value after the labeling of each tilmifloxacin solution, and dividing the difference value of the calculated value and the calculated value by adding the tilmifloxacin standard substance to obtain the condition of the labeling recovery rate.
The specific standard defines the formula as:
the results of the three-electrode system of the electrochemical workstation using the nano copper oxide modified glassy carbon electrode as the working electrode and the measurement and analysis by the DPV method are shown in the table 1, and the recovery rate of the tilmifloxacin with the standard is finally obtained by calculation to be 70.0% -96.6%.
TABLE 1 recovery of temifloxacin
Claims (5)
1. A method for analyzing timixacin by using a modified electrode is characterized by comprising the following steps of:
s40, preparing a tilmicafloxacin mother solution;
s50, preparing a buffer solution;
s60, providing a nano copper oxide modified electrode, wherein the preparation method of the nano copper oxide modified electrode comprises the following steps S10, 20 and 30:
s10, preparing nano copper oxide: adding polyethylene glycol into the copper acetate solution to form a solution A; adding polyethylene glycol into a sodium carbonate solution, dissolving to form a solution B, then uniformly dropwise adding the solution B into the solution A under the ultrasonic condition, performing ultrasonic treatment for a proper time, finally performing magnetic stirring for 30 minutes at 78 ℃ to obtain a black precipitate, centrifuging the black precipitate, repeatedly washing the black precipitate for multiple times, and performing vacuum drying to obtain a copper oxide nano material;
s20, preparing a nano copper oxide suspension: weighing copper oxide nanopowder, placing the copper oxide nanopowder in a sample tube, dripping distilled water into the sample tube to enable the mass ratio of the copper oxide nanopowder to the water to be 1:200, stirring for 5-10 minutes, then adding 1.00mL of 1% PVP into the suspension, and finally placing the sample tube in an ultrasonic cleaner to vibrate and disperse for 30min for later use; shaking for 15-20min before each use so that the nano copper oxide particles can be uniformly dispersed;
s30, modifying glassy carbon electrode: firstly, polishing and grinding a glassy carbon electrode on aluminum oxide powder, then placing the glassy carbon electrode in a beaker filled with dilute sulfuric acid, shaking and cleaning for 5-10min, then taking out and cleaning with distilled water, and finally placing at room temperature for air drying; sucking 10 microliters of the uniformly dispersed nano copper oxide suspension by using a micro-syringe, dripping the suspension on the surface of an electrode core of the glassy carbon electrode by adopting a dripping method, and then drying the electrode core under infrared light to obtain a modified electrode of which the surface is filled with a thin layer of nano copper oxide particles;
s70, adding the tilmicin solution into a buffer solution, using the nano copper oxide modified electrode in the step S60 as a working electrode, a platinum electrode as an auxiliary electrode and a calomel electrode as corresponding reference electrodes to jointly form a three-electrode system, and detecting the tilmicin content by using a cyclic voltammetry.
2. The method of claim 1, wherein in step S50, the buffer is phosphate buffer.
3. The method of claim 2, wherein in step S50, the pH of the phosphate buffer is 7.52.
4. The method of claim 3, wherein in step S70, the nano-copper oxide modified glassy carbon electrode scan rate is 0.10V/S.
5. The method according to claim 1, wherein step S40 specifically comprises the steps of:
accurately weighing a certain mass of tilmicafloxacin technical product, placing the tilmicafloxacin technical product in a beaker, adding water for dissolving, and then fixing the volume to a volumetric flask to form 0.2 × 10-2And (3) placing the tilmicin mother liquor in a dark place for later use.
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