CN111398378B - Preparation method of composite material modified electrode for detecting glucose and electrode - Google Patents
Preparation method of composite material modified electrode for detecting glucose and electrode Download PDFInfo
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Abstract
The invention relates to the field of electrochemical biosensors, and discloses a preparation method of a modified electrode for glucose detection and the electrode. The preparation method of the modified electrode comprises the following steps: firstly, preparing a polymethylene blue modified electrode by adopting a cyclic voltammetry method, then synthesizing nickel oxide nanoflowers by adopting a hydrothermal method, and modifying a dispersion liquid containing nickel oxide on the surface of the polymethylene blue modified electrode by adopting a dripping coating method to construct a nickel oxide/polymethylene blue composite modified electrode. The obtained modified electrode is used as a working electrode, the silver/silver chloride electrode is used as a reference electrode, the platinum electrode is used as a counter electrode to form a three-electrode system, the glucose solution in a certain concentration range is detected by utilizing a chronoamperometry, and the modified electrode has good selectivity and high sensitivity.
Description
Technical Field
The invention relates to a preparation method of an electrochemical biosensor, in particular to a preparation method of a composite material modified electrode for detecting glucose and an electrode.
Background
Diabetes is a chronic disease, seriously harms the health of people and even endangers life. The diagnosis of diabetes can be carried out by detecting the concentration of glucose in human blood, and the health condition of human body can be reflected. Therefore, the realization of the in vitro quantitative detection of the glucose in the blood has important significance for clinical medical diagnosis, the guarantee of human physical and mental health and the like.
The existing methods for detecting glucose comprise spectrophotometry, chromatography, colorimetry, optical rotation method and the like, and have the problems of long detection time, complicated operation steps, expensive equipment and the like. Compared with the methods, the electrochemical analysis method has attracted much attention due to its advantages of high sensitivity, high accuracy, fast response speed, low detection limit, low production cost, and the like.
If the working electrode in the electrochemical analysis method is directly used in the form of a bare electrode without modification, the response signal of glucose is weak, the detection limit is low, and the selectivity of the electrode cannot meet the application requirement in the actual blood sample. Therefore, by utilizing the performance advantages of different materials, more novel composite material modified electrodes aiming at glucose detection are developed, and the sensitivity, detection limit and selectivity of the glucose sensor can be improved.
Disclosure of Invention
The invention aims to provide a preparation method of a composite material modified electrode for detecting glucose and the electrode, aiming at the defects of the prior art, the electrode can be used for quickly and conveniently detecting the glucose, and has the advantages of high sensitivity, low detection limit, good selectivity and higher application value.
The purpose of the invention is realized by the following technical scheme: a composite material modified electrode for detecting glucose is prepared by the following steps:
(1) placing a glassy carbon electrode in a buffer solution with the pH value of 7 and containing 0.8-1.2 mM of methylene blue; carrying out electropolymerization reaction by adopting a cyclic voltammetry, wherein the cyclic potential is-0.4 to +1.6V, the scanning rate is 100mV/s, and the number of cyclic cycles is 15 cycles to prepare a polymethylene blue modified electrode;
(2) dissolving 0.75-1.0 g of sodium dodecyl sulfate in 80ml of deionized water, and adding 0.75-1.1 g of nickel chloride hexahydrate and 0.50-0.75 g of urea under continuous stirring; transferring the obtained mixed solution to a 100 ml Teflon stainless steel autoclave, reacting for 6-10 h at 140-180 ℃, and cooling to room temperature; collecting the generated light green precipitate, centrifuging at 8000r/min for 10min, and washing with deionized water and ethanol for several times; placing the obtained product in a vacuum drying oven, keeping the temperature of 40-70 ℃ and drying for 9-12 h to prepare nickel oxide nanoflowers;
(3) adding 10-20 mg of nickel oxide powder prepared in the step into 2ml of dimethylformamide solution containing 5% of perfluorosulfonic acid resin, and carrying out magnetic stirring at room temperature; absorbing 2 mu L of light green suspension liquid drop on the surface of the polymethylene blue modified electrode by using a liquid transfer gun, placing the electrode under infrared light for drying for 40min at a position of 20-25 cm, and repeating the modification step of the electrode for 3 times; and washing the electrode with deionized water and drying in the air to obtain the nickel oxide/polymethylene blue modified electrode.
The electrolyte in the step (1) is a buffer solution which contains 1mM of methylene blue and has the pH value of 7; the scanning rate during the electropolymerization is 100mV/s, and the number of cycles is 15.
Dissolving 0.86g of sodium dodecyl sulfate in 80ml of deionized water, and then adding 0.95g of nickel chloride hexahydrate and 0.60g of urea; reacting the mixed solution at the temperature of 160 ℃ for 8 hours at high temperature and high pressure; the resulting precipitate was centrifuged at 8000r/min for 10 minutes and, after washing, dried in vacuo at 60 ℃ for 11 h.
Mixing the 15mg nickel oxide powder with 2ml of dimethylformamide solution containing perfluorosulfonic acid resin in the step (3); and (3) sucking 2 mu L of light green suspension liquid drop on the surface of the polymethylene blue modified electrode by using a liquid transfer gun, and drying for 40min under infrared light.
The nickel oxide/polymethylene blue composite material-based modified electrode prepared by the preparation method.
The invention has the beneficial effects that:
(1) the invention provides a composite material modified electrode for detecting glucose, which combines high-conductivity polymethylene blue and nickel oxide with a catalytic effect on glucose to prepare a nickel oxide/polymethylene blue composite material modified electrode. The prepared modified electrode can realize the quantitative detection of the glucose concentration, and the catalytic performance and the detection effect of the modified electrode are superior to those of a bare electrode or an electrode modified by a single material;
(2) the glucose electrochemical sensor constructed by the invention has the advantages of simple preparation method, high detection speed, low cost, high sensitivity and good selectivity, and can be used for detecting actual samples.
Drawings
FIG. 1 is a cyclic voltammogram during the preparation of a polymethylene blue modified electrode;
FIG. 2 shows a bare electrode (a) and a polymethylene blue modified electrode (b) at 1mmol/L of Fe (CN)6 3-/4-Electrochemical response results in a solution (containing 0.1mol/L KCl);
FIG. 3 is an XRD pattern of nickel oxide;
FIG. 4 is a scanning electron micrograph of nickel oxide;
FIG. 5 is a graph showing the results of cyclic voltammetry scans of a bare electrode, a nickel oxide-modified electrode, and a nickel oxide/polymethylene blue-modified electrode in a solution containing 0.1mmol/L glucose;
FIG. 6 shows the glucose concentration of the nickel oxide/polymethylene blue modified electrode at 3X 10-6~8×10-5In the mol/L range, the current responds to the change chart along with time;
FIG. 7 is a standard curve of glucose response current versus concentration gradient on a nickel oxide/polymethylene blue modified electrode;
fig. 8 is an anti-interference test curve of the nickel oxide/polymethylene blue modified electrode.
Detailed Description
The present invention will be described in further detail with reference to specific examples, but the embodiments of the present invention are not limited thereto.
Example 1
The polymethylene blue composite material modified electrode is prepared by the following method:
(1) respectively mixing 0.5 mu m and 0.05 mu m aluminum oxide powder with deionized water and a volume ratio of 1:1, pouring a dispersion liquid formed by 0.5 mu m aluminum oxide on a rough chamois, polishing a glassy carbon electrode, pouring a dispersion liquid formed by 0.05 mu m aluminum oxide on a fine chamois, and continuously polishing the electrode to a mirror surface. Ultrasonically cleaning the polished electrode with dilute nitric acid, absolute ethyl alcohol and deionized water for 3 times for 30s each time;
(2) placing the cleaned electrode in 0.5mol/L dilute sulfuric acid solution, performing electrochemical activation by adopting cyclic voltammetry, wherein the potential interval is-0.5V to +1.2V, the scanning rate is 100mV/s, and the number of cycles is 50, then cleaning the electrode by using deionized water, and airing;
(3) placing the dried electrode at 1mmol/L K3[Fe(CN)6]Cyclic voltammetry is carried out in the solution (containing 0.1mol/L KCl), the potential interval is-0.06V to +0.6V, and the sweep rate is 50 mV/s. When [ Fe (CN)6]3-/4-The difference of the oxidation-reduction peak potentials on the electrodes is about 68mV, which indicates that the surface of the glassy carbon electrode has reached the requirement of cleaning, the reversibility of the electrode is good, and then the electrode is cleaned by deionized water and dried for standby;
(4) and (3) placing the pretreated glassy carbon electrode in a PBS (phosphate buffer solution) containing 1mM of methylene blue and having a pH value of 7, and performing electropolymerization reaction by adopting a cyclic voltammetry, wherein the cyclic potential is-0.4 to +1.6V, the scanning rate is 100mV/s, and the number of cyclic cycles is 15 cycles to prepare a polymethylene blue modified electrode, and the cyclic voltammetry curve in the preparation process is shown in figure 1.
The electrochemical characterization of the polymethylene blue modified electrode of the embodiment comprises the following specific operation steps:
(1) taking a polymethylene blue composite material modified electrode as a working electrode, a silver electrode or a silver chloride electrode as a reference electrode, and a platinum electrode as a counter electrode, constructing a three-electrode system, and connecting the three-electrode system with an electrochemical workstation;
(2) 1mM Fe (CN)6 3-/4-Pouring the solution (containing 0.1M KCl) into an electrolytic cell;
(3) and (3) performing cyclic voltammetry test on the composite material, wherein the potential interval is-0.6V to +0.6V, and the sweep rate is 50 mV/s. Fig. 2 shows the response of the bare electrode and the polymethylene blue modified electrode prepared in the above step in the electrochemical test under the same conditions, and the result shows that the polymethylene blue modified electrode can generate a larger response current and has stronger conductivity, and the polymethylene blue material has better electron transmission capability.
Example 2
The difference between the preparation method of the polymethylene blue composite material modified electrode described in this embodiment and embodiment 1 is that: in the step (4), the pretreated glassy carbon electrode is placed in PBS (phosphate buffer solution) containing 0.8mM or 1.2mM of methylene blue and having a pH value of 7, and electropolymerization reaction is carried out by adopting a cyclic voltammetry, wherein the cyclic potential is-0.4 to +1.6V, the scanning rate is 100mV/s, and the number of cyclic turns is 15, so that the polymethylene blue modified electrode is prepared.
Example 3
The nickel oxide/polymethylene blue composite material modified electrode is prepared by the following method:
(1) 0.86g of sodium dodecyl sulfate was weighed into a beaker, dissolved in 80ml of deionized water, and 0.95g of nickel chloride hexahydrate and 0.60g of urea were added with vigorous stirring, and then the light green mixed solution was transferred to a 100 ml teflon stainless steel autoclave and placed in an oven to react for 8 hours at 160 ℃. After naturally cooling to room temperature, the waste liquid is poured out, and light green precipitates are collected and centrifuged for 10min at the rotating speed of 8000 r/min. The obtained precipitate was repeatedly washed with deionized water and then with ethanol. And (3) putting the cleaned precipitate into a vacuum drying oven, and keeping the temperature of 60 ℃ for drying for 11h to obtain the nickel oxide nanoflower. FIG. 3 is an XRD diagram of nickel oxide prepared by the present invention, the product is NiO, and the purity of the prepared product is higher; FIG. 4 is a scanning electron microscope image of nickel oxide prepared by the present invention, and the product is nickel oxide nanoflower with relatively uniform size and larger specific surface area;
(2) and (3) adding 15mg of nickel oxide powder prepared in the step into 2mL of dimethylformamide solution containing 5% of perfluorosulfonic acid resin, and magnetically stirring at room temperature for 2 h. And (3) sucking 2 mu L of light green suspension liquid by using a liquid transfer gun to drop on the surface of the polymethylene blue modified electrode prepared in the embodiment 1, drying the electrode at a position of 25cm under infrared light for 40min, repeating the step after the surface of the electrode is fully dried, and dripping 3 times of nickel oxide-dimethylformamide dispersion liquid to prepare the nickel oxide/polymethylene blue modified electrode.
Example 4
The difference between the preparation method of the nickel oxide/polymethylene blue composite material modified electrode in this embodiment and embodiment 3 is that: in the step (1), 0.50g or 1.0g of sodium dodecyl sulfate is placed in a beaker, dissolved with deionized water, and then 0.95g of nickel chloride hexahydrate and 0.60g of urea are added for reaction at high temperature and high pressure. The obtained precipitate was centrifuged and washed repeatedly with deionized water and ethanol. And (3) putting the cleaned precipitate into a vacuum drying oven, and keeping the temperature of 60 ℃ for drying for 11h to obtain the nickel oxide nanoflower.
Example 5
The difference between the preparation method of the nickel oxide/polymethylene blue composite material modified electrode in this embodiment and embodiment 3 is that: in the step (2), 0.86g of sodium dodecyl sulfate is placed in a beaker, dissolved by deionized water, and then 0.75g or 1.1g of nickel chloride hexahydrate and 0.60g of urea are added for reaction at high temperature and high pressure. The obtained precipitate was centrifuged and washed repeatedly with deionized water and ethanol. And (4) putting the cleaned precipitate into a vacuum drying oven for drying to obtain the nickel oxide nanoflower.
Example 6
The difference between the preparation method of the nickel oxide/polymethylene blue composite material modified electrode in this embodiment and embodiment 3 is that: in the step (2), 0.86g of sodium dodecyl sulfate is dissolved in 80ml of deionized water, and then 0.95g of nickel chloride hexahydrate and 0.50g or 0.75g of urea are added to react at high temperature and high pressure. And centrifuging the obtained precipitate, repeatedly washing with deionized water and ethanol, and drying to obtain the nickel oxide nanoflower.
Example 7
The modified electrode is used for feasibility verification of glucose concentration, and comprises the following specific operation steps:
(1) taking the nickel oxide/polymethylene blue composite material modified electrode prepared in the embodiment 2 as a working electrode, a silver electrode or a silver chloride electrode as a reference electrode, and a platinum electrode as a counter electrode, constructing a three-electrode system, and connecting the three-electrode system with an electrochemical workstation;
(2) adding 0.1mmol/L grape solution into an electrolytic cell;
(3) scanning in a potential interval of +0.3V to +0.7V by adopting a cyclic voltammetry at a scanning speed of 120 mV/s. The obtained cyclic voltammogram was compared with the bare electrode and the nickel oxide-modified electrode, and the result is shown in fig. 5. Compared with a bare electrode and a nickel oxide modified electrode, the modified electrode has an obvious response peak to glucose, and shows that the nickel oxide/polymethylene blue composite material plays a role in electrocatalysis of glucose oxidation reaction, so that the reaction current is enhanced and the reaction potential is reduced.
Example 8
The modified electrode is used for measuring the concentration of glucose, and comprises the following specific operation steps:
(1) taking the nickel oxide/polymethylene blue composite material modified electrode prepared in the embodiment 2 as a working electrode, a silver electrode or a silver chloride electrode as a reference electrode, and a platinum electrode as a counter electrode, constructing a three-electrode system, and connecting the three-electrode system with an electrochemical workstation;
(2) preparing 0.1mol/L NaOH solution, pouring the NaOH solution into an electrolytic cell, and inserting a three-electrode system into the electrolytic cell;
(3) setting the working mode of the electrochemical workstation as a chronoamperometry method, setting the potential to be + 0.47V and the sampling interval to be 0.1s, dropwise adding 1mL of glucose solution with the concentration gradient of 0.1mmol/L, 0.25mmol/L, 0.50mmol/L, 0.75mmol/L and 1mmol/L into the electrolytic cell every 50s after the current is stabilized for about 150s, and continuously dropwise adding each gradient solution for 4 times for a total of 200s to obtain the relation between the glucose response current and the time. The obtained result is shown in fig. 6, with each addition of the glucose solution, the glucose concentration of the whole solution system in the electrolytic cell changes, the electrochemical response signal generated by the glucose concentration changes, and the change value increases with the increase of the concentration of the dropwise added glucose solution;
(4) fitting the obtained response current with the dropwise added glucose concentration according to the result obtained in the step (3), and drawing a standard curve by using origin software, as shown in fig. 7. If the glucose concentration is 3X 10-6 ~ 5×10-5In the mol/L range, the current change value during measurement is substituted into the formula i = -0.0292c-0.0783,obtaining the concentration of the glucose solution; if the glucose concentration is 5X 10-5~8×10-5In the mol/L range, the current change value during measurement is substituted into the formula i = -0.0832c + 2.8925, and the concentration of the glucose solution is obtained. Wherein the concentration c is in units of μmol/L and the current is in units of μ A;
(5) setting the potential to be + 0.47V in a timing current method mode, dropwise adding 1mL of glucose solution with the concentration gradient of 0.5mmol/L into the electrolytic cell to obtain a current response change value, substituting the peak current value into a formula i = -0.0292c-0.0783, and calculating the deviation of the obtained concentration and the actual concentration to be less than 2%.
Example 9
The anti-interference capability of the modified electrode prepared by the invention is tested by a time-lapse current method, and the specific operation steps are as follows:
(1) taking the nickel oxide/polymethylene blue composite material modified electrode prepared in the embodiment 2 as a working electrode, a silver electrode or a silver chloride electrode as a reference electrode, and a platinum electrode as a counter electrode, constructing a three-electrode system, and connecting the three-electrode system with an electrochemical workstation;
(2) preparing a glucose solution with the concentration of 0.1mmol/L and uric acid, ascorbic acid and a solution with the concentration of 10mmol/L, and inserting a three-electrode system into an electrolytic cell;
(3) the working mode of the electrochemical workstation is set as a chronoamperometry method, the potential is + 0.47V, the sampling interval is 0.1s, glucose, uric acid and ascorbic acid solution are added into the electrolytic cell in an unordered manner every 100s after the current is stabilized for about 250s, the relation between the response current and the time is obtained, and the obtained result is shown in FIG. 8. The response current of the modified electrode is hardly influenced by 100 times of uric acid and ascorbic acid, which shows that the nickel oxide/polymethylene blue composite material modified electrode has good anti-interference capability.
The above-described embodiments are intended to illustrate rather than to limit the invention, and any modifications and variations of the present invention are within the spirit of the invention and the scope of the appended claims.
Claims (4)
1. A preparation method of a composite material modified electrode for detecting glucose is characterized by comprising the following steps:
(1) placing a glassy carbon electrode in an electrolyte, wherein the electrolyte is a buffer solution containing 0.8-1.2 mM of methylene blue and having a pH value of 7; carrying out electropolymerization reaction by adopting a cyclic voltammetry, wherein the cyclic potential is-0.4 to +1.6V, the scanning rate is 100mV/s, and the number of cyclic cycles is 15 cycles to prepare a polymethylene blue modified electrode;
(2) dissolving 0.75-1.0 g of sodium dodecyl sulfate, 0.75-1.1 g of nickel chloride hexahydrate and 0.50-0.75 g of urea in deionized water; transferring the obtained mixed solution into a Teflon stainless steel autoclave, and reacting for 6-10 h at 140-180 ℃ to obtain nickel oxide nanoflowers;
(3) taking 10-20 mg of nickel oxide powder prepared in the step (2), adding the powder into 2ml of dimethylformamide solution containing 5% of perfluorinated sulfonic acid resin to obtain light green suspension, dripping the light green suspension on the surface of the polymethylene blue modified electrode prepared in the step (1), and drying the electrode at a position of 20-25 cm under infrared light; washing the electrode with deionized water and drying in the air to obtain a nickel oxide/polymethylene blue modified electrode;
in the step (1), the electrolyte is a buffer solution containing 1mM of methylene blue and having a pH value of 7; the scanning rate during the electropolymerization is 100mV/s, and the number of cycles is 15.
2. The method of claim 1, wherein: in the step (2), 0.86g of sodium dodecyl sulfate is dissolved in 80ml of deionized water, and 0.95g of nickel chloride hexahydrate and 0.60g of urea are added under continuous stirring; reacting the mixed solution at the high temperature of 160 ℃ for 8 hours under high pressure; the resulting precipitate was centrifuged at 8000r/min for 10min and after washing was dried in vacuo at 60 ℃ for 11 h.
3. The method of claim 1, wherein: in the step (3), 15mg of nickel oxide powder is mixed with 2ml of dimethylformamide solution containing perfluorosulfonic acid resin; the following electrode modification steps were repeated three times: and (3) sucking 2 mu L of light green suspension liquid drop on the surface of the polymethylene blue modified electrode by using a liquid transfer gun, and drying for 40min under infrared light.
4. The modified electrode based on the nickel oxide/polymethylene blue composite material obtained by the preparation method of claim 1.
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