CN112649482A - Enzyme-free electrochemical urea sensor based on Ni-MOFs composite material and preparation method and application thereof - Google Patents
Enzyme-free electrochemical urea sensor based on Ni-MOFs composite material and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides an enzyme-free electrochemical urea sensor based on a Ni-MOFs composite material and a preparation method and application thereof. The sensor prepared by the invention has excellent performances including higher sensitivity, faster response speed, wider detection range and good anti-interference performance.
Description
Technical Field
The invention relates to the technical field of electrochemical biosensors, in particular to an enzyme-free electrochemical urea sensor based on a Ni-MOFs composite material, and a preparation method and application thereof.
Background
Urea is produced by the liver, then fuses into the blood and is finally excreted by the urine through the kidneys, a small amount of urea is also excreted through sweat. Since abnormalities in urea levels in humans are often associated with a variety of diseases such as cardiac metabolic disorders, chronic kidney disease, and renal failure, detection of urea levels plays a crucial role in assessing various metabolic disorders; in addition, some merchants mix urea in milk and feed to improve the detection content of protein, but too much urea in human bodies can cause harm to livers, kidneys and alveoli of the human bodies due to exceeding the absorption capacity of the human bodies, so the detection of urea has important significance in food science; in addition, urea is also a high-concentration nitrogen fertilizer and is used in the agricultural field, and pesticide residues of vegetables are also related to urea detection; in order to prevent the frequency of changing the swimming pool water from being reduced for reducing the cost of a merchant and further to prevent the damage to the body of a customer, the urea content detection in the swimming pool water also has important application prospect.
The detection methods of urea include colorimetry, spectrometry, chromatography, etc., each of which has its own advantages, but continuous rapid detection of urea cannot be achieved, and thus electrochemical biosensors having rapid response and high sensitivity have been widely researched and rapidly developed.
The electrochemical sensor converts a signal generated by the reaction of the identification original and the urea into a digital electric signal by taking the modified material on the electrode as the identification original, so that the concentration of the urea is calculated according to the signal intensity. According to different recognition elements, namely different catalysis principles, the electrochemical urea sensor can be divided into an enzyme-based electrochemical urea sensor and an enzyme-free electrochemical urea sensor.
The enzyme-based urea electrochemical sensor detects the concentration of urea by utilizing the specific electrochemical reaction of urease and urea and converting the specific electrochemical reaction into an electric signal. The enzyme-based electrochemical urea sensor has the advantages of good specificity and high sensitivity, but the enzyme price is high, the fixing process is complex, and the main defect of poor stability of the enzyme is still difficult to overcome due to the biological essential attribute of the enzyme; in addition, the enzyme has limited working conditions, and heavy metal ions, magnetic nanoparticles and heat can inactivate the enzyme, thereby preventing the application of the enzyme sensor.
The catalysis principle of the enzyme-free urea electrochemical sensor is that urea generates oxidation/reduction reaction on a proper electrode, and the catalysis material generates electrocatalytic oxidation on the urea, so that the oxidation current is increased, and the detected oxidation peak current and the urea concentration have a linear relation. The current type electrochemical urea sensor changes the urea concentration under a fixed potential, measures the current to obtain the functional relation between the current and the urea concentration, and then calculates the unknown urea concentration according to the current.
The non-enzyme electrochemical urea sensor has the advantages of low cost, simple preparation process and good repeatability and stability. The research of the non-enzyme electrochemical urea sensor mainly focuses on improving the sensitivity and the selectivity of the non-enzyme electrochemical urea sensor. The sensitivity is greatly improved by researches such as preparing catalytic materials with better catalytic performance, such as metal nano materials, composite nano materials and the like, and improving the specific surface area of the catalytic materials. On the other hand, although the interference of biomolecules such as dopamine, glucose, ascorbic acid and the like is reduced by regulating the working potential of the urea sensor, the selectivity of the enzyme-free urea sensor is still insufficient compared with the specific catalysis of the enzyme-based urea sensor.
The existing nickel-based enzyme-free urea sensor has been widely researched, but the following problems still exist: the sensitivity of the constructed urea sensor is not high enough; the sensor has complex preparation process and harsh experimental conditions, and is not easy to realize industrial production.
Disclosure of Invention
The invention overcomes the defects in the prior art, and provides an enzyme-free electrochemical urea sensor based on a Ni-MOFs composite material, and a preparation method and application thereof.
The purpose of the invention is realized by the following technical scheme.
An enzyme-free electrochemical urea sensor based on Ni-MOFs composite material and a preparation method thereof are disclosed, wherein the enzyme-free electrochemical urea sensor comprises a reference electrode, a counter electrode and a modified working electrode, the modified working electrode comprises a working electrode and an active catalytic material solidified on the surface of the working electrode, the reference electrode adopts an Ag/AgCl reference electrode, the counter electrode adopts a platinum wire electrode, the working electrode adopts a gold electrode, the active catalytic material is Ni-MOFs composite material, and the modified working electrode is prepared according to the following steps:
and 2, dropwise adding the Ni-MOF dispersion liquid prepared in the step 1 on the surface of the gold electrode, standing in the air for 1-3h, and obtaining the Ni-MOFs composite material modified gold electrode, namely the modified working electrode, wherein the usage amount of the Ni-MOF dispersion liquid is 6-10 mu L.
In the step 1, the mass concentration of the Ni-MOF composite material in the Ni-MOF dispersion liquid is 4-5 mg/mL.
In the step 2, the Ni-MOF composite material is prepared by adopting a one-step hydrothermal method according to the following steps: nickel nitrate hexahydrate (Ni (NO)3)2·6H2O) was dissolved in methanol, and 1,3, 5-tribenzoic acid (H) was added to the above mixed solution3BTC) under the room temperature of 20-25 ℃ and is subjected to ultrasonic treatment to be uniformly mixed, the solution is placed at the temperature of 120-200 ℃, after 24-30h of reaction, centrifugation, methanol cleaning and drying are carried out, and the Ni-MOF composite material is obtained, wherein nickel nitrate hexahydrate (Ni (NO)3)2·6H2O) and 1,3, 5-Tribenzoic acid (H)3BTC) is (1-3) to (3-1).
Nickel nitrate hexahydrate (Ni (NO)3)2·6H2O) and 1,3, 5-Tribenzoic acid (H)3BTC) is 1:2-2:1, the reaction temperature is 140-.
In step 2, the gold electrode needs to be pretreated, and the pretreatment process comprises the following steps: applying gold electrode to Al2O3Grinding the polishing powder in an 8-shaped manner, performing ultrasonic treatment in deionized water for 0.5h, and blow-drying the surface of the polishing powder by using nitrogen gas to obtain the gold electrode.
In step 2, the amount of Ni-MOF dispersion was 6-8. mu.L.
The invention has the beneficial effects that: the sensor prepared by the invention has excellent performances including higher sensitivity, faster response speed, wider detection range and good anti-interference performance.
Drawings
FIG. 1 is a Scanning Electron Microscope (SEM) photograph of Ni-MOFs composite material prepared by the invention under different magnifications;
FIG. 2 is a Cyclic Voltammetry (CV) curve measured by an enzyme-free electrochemical urea sensor based on Ni-MOFs composite material, prepared by the method, under different urea concentrations;
FIG. 3 is a time-current (i-t) curve and a corresponding linear fitting curve measured by the enzyme-free electrochemical urea sensor based on the Ni-MOFs composite material and prepared by the invention under the condition of changing the urea concentration;
FIG. 4 is a time-current (i-t) curve of the enzyme-free electrochemical urea sensor based on the Ni-MOFs composite material, which is prepared by the invention, when various interfering substances are added.
Detailed Description
The technical solution of the present invention is further illustrated by the following specific examples.
Example 1
(1) Preparation of Ni-MOFs composite material
A simple one-step hydrothermal method was used, as follows: 4.4mmol of Ni (NO)3)2·6H2O was dissolved in methanol and then an equimolar amount of H was added3BTC, stirring at room temperature and carrying out ultrasonic treatment for 1h respectively to uniformly mix the solution, and then carrying out hydrothermal treatment on the solution at 150 ℃ for 24 h. After the reaction is finished, centrifuging to collect the product, repeatedly washing the product with methanol for several times, and drying the obtained product in a vacuum oven at 60 ℃ for 24 hours for later use. The SEM photograph of the prepared Ni-MOFs composite material is shown in the attached figure 1.
(2) Modified working electrode
Applying gold electrode to Al2O3Polishing the polishing powder to form an 8-shaped surfacePerforming ultrasonic treatment in ionized water for 0.5h, and then drying the surface of the ionized water by using nitrogen to obtain a gold electrode for later use; weighing 5mg of prepared Ni-MOF material, dispersing in 1mL of deionized water, stirring and carrying out ultrasonic treatment for 1 hour respectively to fully disperse the Ni-MOF material; and sucking 8 mu of LNi-MO dispersion liquid by using a liquid-transferring gun, dripping the dispersion liquid on the surface of the ground gold electrode, standing the gold electrode in the air for 2 hours to obtain the Ni-MOF modified gold electrode which is used as a working electrode.
(3) Construction of an electrochemical Urea sensor
The Ni-MOFs modified gold electrode, a counter electrode and a reference electrode form a three-electrode system (a platinum wire electrode is used as the counter electrode, and an Ag/AgCl reference electrode is used as the reference electrode) to construct the urea sensor
The electrochemical workstation was opened and the electrochemical test was performed at room temperature in 0.1M NaOH solution using cyclic voltammetry at a voltage range of 0-0.8V and a scan rate of 50 mV/s. And (4) dropwise adding a certain amount of urea solution in sequence, uniformly stirring, and then testing.
The cyclic voltammogram of the electrochemical urea sensor based on the Ni-MOFs composite material prepared in the embodiment after urea with different concentrations is added is shown in FIG. 2. The oxidation peak-to-peak current increased with increasing urea concentration, indicating that the Ni-MOF material has electrocatalytic oxidation activity on urea.
Example 2
(1) Preparation of Ni-MOFs composite material
A simple one-step hydrothermal method was used, as follows: 4.4mmol of Ni (NO)3)2·6H2O was dissolved in methanol and 6mmol H was added3BTC, stirring at room temperature and carrying out ultrasonic treatment for 1h respectively to uniformly mix the solution, and then carrying out hydrothermal treatment on the solution at 150 ℃ for 28 h. After the reaction is finished, centrifuging to collect the product, repeatedly washing the product with methanol for several times, and drying the obtained product in a vacuum oven at 60 ℃ for 24 hours for later use.
(2) Modified working electrode
Applying gold electrode to Al2O3Grinding the polishing powder in an 8-shaped manner, performing ultrasonic treatment in deionized water for 0.5h, and blow-drying the surface of the polishing powder by using nitrogen to obtain a gold electrode for later use; weighing 5mg of prepared Ni-MOF material, and dividingDispersing in 1mL of deionized water, stirring and performing ultrasonic treatment for 1 hour respectively to fully disperse the deionized water; and sucking 8 mu of LNi-MO dispersion liquid by using a liquid-transferring gun, dripping the dispersion liquid on the surface of the ground gold electrode, standing the gold electrode in the air for 2 hours to obtain the Ni-MOF modified gold electrode which is used as a working electrode.
(3) Construction of an electrochemical Urea sensor
The Ni-MOFs modified gold electrode, a counter electrode and a reference electrode form a three-electrode system (a platinum wire electrode is used as the counter electrode, and an Ag/AgCl reference electrode is used as the reference electrode) to construct the urea sensor
Opening an electrochemical workstation, carrying out an electrochemical test at room temperature in a 0.1M NaOH solution, adopting a timing current method, wherein the working voltage is 0.5V, continuously adding quantitative urea in the test process and stirring the urea at the rotating speed of 100 revolutions per minute.
The electrochemical urea sensor based on the Ni-MOFs composite material prepared in the embodiment is characterized in that urea is continuously added dropwise into a 0.1M NaOH solution, the current is increased in a stepwise manner, an i-t curve is obtained, and a current-concentration calibration curve is drawn, and the result is shown in the attached figure 3. With the continuous injection of urea, the current response increases sharply, the inset is an enlargement of the i-t curve, and it can be seen that the response time of the current after the addition of urea is 4-5 s. The electrode showed good catalytic response in the range of 0.1-6mM with sensitivity of 152.5. mu.A mM at low concentrations of 0.1mM to 2mM-1Linear correlation coefficient of 0.998, sensitivity of 92.7. mu.A mM at high concentrations of 1 to 6mM-1And a linear correlation coefficient of 0.993.
Example 3
(1) Preparation of Ni-MOFs composite material
A simple one-step hydrothermal method was used, as follows: 4.4mmol of Ni (NO)3)2·6H2O was dissolved in methanol and then an equimolar amount of H was added3BTC, stirring at room temperature and carrying out ultrasonic treatment for 1h respectively to uniformly mix the solution, and then carrying out hydrothermal treatment on the solution at 150 ℃ for 28 h. After the reaction is finished, centrifuging to collect the product, repeatedly washing the product with methanol for several times, and drying the obtained product in a vacuum oven at 60 ℃ for 24 hours for later use.
(2) Modified working electrode
Applying gold electrode to Al2O3Grinding the polishing powder in an 8-shaped manner, performing ultrasonic treatment in deionized water for 0.5h, and blow-drying the surface of the polishing powder by using nitrogen to obtain a gold electrode for later use; weighing 5mg of prepared Ni-MOF material, dispersing in 1mL of deionized water, stirring and carrying out ultrasonic treatment for 1 hour respectively to fully disperse the Ni-MOF material; and sucking 8 mu of LNi-MO dispersion liquid by using a liquid-transferring gun, dripping the dispersion liquid on the surface of the ground gold electrode, standing the gold electrode in the air for 2 hours to obtain the Ni-MOF modified gold electrode which is used as a working electrode.
(3) Construction of an electrochemical Urea sensor
The Ni-MOFs modified gold electrode, a counter electrode and a reference electrode form a three-electrode system (a platinum wire electrode is used as the counter electrode, and an Ag/AgCl reference electrode is used as the reference electrode) to construct the urea sensor
Open the electrochemical workstation, perform electrochemical tests at room temperature, perform the i-t test in 0.1M NaOH solution in the presence of various interferents (including creatinine, ascorbic acid, glucose and uric acid), at an operating voltage of 0.5V, at a concentration similar to that present in a normal urine sample, and the results are shown in FIG. 4
Compared with the current response when the urea is added, the electrochemical urea sensor based on the Ni-MOFs composite material prepared in the embodiment has only slight disturbance of the current when various interferents are added, and the fact that the addition of each interferent does not have considerable interference in the urea detection process means that the Ni-MOF electrode is suitable for detecting the urea level in an actual sample.
The invention has been described in an illustrative manner, and it is to be understood that any simple variations, modifications or other equivalent changes which can be made by one skilled in the art without departing from the spirit of the invention fall within the scope of the invention.
Claims (10)
1. An enzyme-free electrochemical urea sensor based on a Ni-MOFs composite material is characterized in that: the modified working electrode comprises a working electrode and an active catalytic material solidified on the surface of the working electrode, wherein the reference electrode adopts an Ag/AgCl reference electrode, the counter electrode adopts a platinum wire electrode, the working electrode adopts a gold electrode, the active catalytic material is a Ni-MOFs composite material, and the modified working electrode is carried out according to the following steps:
step 1, dispersing a Ni-MOF composite material in deionized water, stirring and performing ultrasonic treatment to fully disperse the Ni-MOF composite material to obtain a Ni-MOF dispersion liquid, wherein the mass concentration of the Ni-MOF composite material in the Ni-MOF dispersion liquid is 3-6 mg/mL;
and 2, dropwise adding the Ni-MOF dispersion liquid prepared in the step 1 on the surface of the gold electrode, standing in the air for 1-3h, and obtaining the Ni-MOFs composite material modified gold electrode, namely the modified working electrode, wherein the usage amount of the Ni-MOF dispersion liquid is 6-10 mu L.
2. The enzyme-free electrochemical urea sensor based on Ni-MOFs composite material according to claim 1, wherein: in the step 1, the mass concentration of the Ni-MOF composite material in the Ni-MOF dispersion liquid is 4-5 mg/mL.
3. The enzyme-free electrochemical urea sensor based on Ni-MOFs composite material according to claim 1, wherein: in the step 2, the Ni-MOF composite material is prepared by adopting a one-step hydrothermal method according to the following steps: nickel nitrate hexahydrate (Ni (NO)3)2·6H2O) was dissolved in methanol, and 1,3, 5-tribenzoic acid (H) was added to the above mixed solution3BTC) under the room temperature of 20-25 ℃ and is subjected to ultrasonic treatment to be uniformly mixed, the solution is placed at the temperature of 120-200 ℃, after 24-30h of reaction, centrifugation, methanol cleaning and drying are carried out, and the Ni-MOF composite material is obtained, wherein nickel nitrate hexahydrate (Ni (NO)3)2·6H2O) and 1,3, 5-Tribenzoic acid (H)3BTC) in a molar ratio of (1-3) to (3-1), preferably, nickel nitrate hexahydrate (Ni (NO)3)2·6H2O) and 1,3, 5-Tribenzoic acid (H)3BTC) is 1:2-2:1, the reaction temperature is 140-.
4. The enzyme-free electrochemical urea sensor based on Ni-MOFs composite material according to claim 1, wherein: in step 2, the gold electrode needs to be pretreated, and the pretreatment process comprises the following steps: applying gold electrode to Al2O3Grinding the polishing powder into an 8-shaped surface, performing ultrasonic treatment in deionized water for 0.5h, and blow-drying the surface of the polishing powder by using nitrogen to obtain the gold electrode, wherein the usage amount of the Ni-MOF dispersion liquid is 6-8 mu L.
5. The preparation method of the non-enzyme electrochemical urea sensor based on the Ni-MOFs composite material is characterized by comprising the following steps of: the modified working electrode comprises a working electrode and an active catalytic material solidified on the surface of the working electrode, wherein the reference electrode adopts an Ag/AgCl reference electrode, the counter electrode adopts a platinum wire electrode, the working electrode adopts a gold electrode, the active catalytic material is a Ni-MOFs composite material, and the modified working electrode is carried out according to the following steps:
step 1, dispersing a Ni-MOF composite material in deionized water, stirring and performing ultrasonic treatment to fully disperse the Ni-MOF composite material to obtain a Ni-MOF dispersion liquid, wherein the mass concentration of the Ni-MOF composite material in the Ni-MOF dispersion liquid is 3-6 mg/mL;
and 2, dropwise adding the Ni-MOF dispersion liquid prepared in the step 1 on the surface of the gold electrode, standing in the air for 1-3h, and obtaining the Ni-MOFs composite material modified gold electrode, namely the modified working electrode, wherein the usage amount of the Ni-MOF dispersion liquid is 6-10 mu L.
6. The method for preparing the enzyme-free electrochemical urea sensor based on the Ni-MOFs composite material, according to claim 1, wherein: in the step 1, the mass concentration of the Ni-MOF composite material in the Ni-MOF dispersion liquid is 4-5 mg/mL.
7. The method for preparing the enzyme-free electrochemical urea sensor based on the Ni-MOFs composite material, according to claim 1, wherein: in the step 2, the Ni-MOF composite material is prepared by adopting a one-step hydrothermal method according to the following steps: will sixHydrated nickel nitrate (Ni (NO)3)2·6H2O) was dissolved in methanol, and 1,3, 5-tribenzoic acid (H) was added to the above mixed solution3BTC) under the room temperature of 20-25 ℃ and is subjected to ultrasonic treatment to be uniformly mixed, the solution is placed at the temperature of 120-200 ℃, after 24-30h of reaction, centrifugation, methanol cleaning and drying are carried out, and the Ni-MOF composite material is obtained, wherein nickel nitrate hexahydrate (Ni (NO)3)2·6H2O) and 1,3, 5-Tribenzoic acid (H)3BTC) in a molar ratio of (1-3) to (3-1), preferably, nickel nitrate hexahydrate (Ni (NO)3)2·6H2O) and 1,3, 5-Tribenzoic acid (H)3BTC) is 1:2-2:1, the reaction temperature is 140-.
8. The method for preparing the enzyme-free electrochemical urea sensor based on the Ni-MOFs composite material, according to claim 1, wherein: in step 2, the gold electrode needs to be pretreated, and the pretreatment process comprises the following steps: applying gold electrode to Al2O3Grinding the polishing powder into an 8-shaped surface, performing ultrasonic treatment in deionized water for 0.5h, and blow-drying the surface of the polishing powder by using nitrogen to obtain the gold electrode, wherein the usage amount of the Ni-MOF dispersion liquid is 6-8 mu L.
9. Use of the enzyme-free electrochemical urea sensor based on Ni-MOFs composite material according to any one of claims 1 to 4 for the detection of urea molecules.
10. Use according to claim 9, characterized in that: the response time of current after the current is 4-5s after the current is added into the non-enzyme electrochemical urea sensor based on the Ni-MOFs composite material, the modified working electrode shows good catalytic response in the range of 0.1-6mM, and the linear curve is that y is 0.0036+0.1525x, R is 0.1-2mM under low concentration20.998, sensitivity 152.5 μ A mM-1At high concentrations of 1-6mM, the linear curve is y-0.1255 +0.0927x, R20.993, sensitivity 92.7 μ A mM-1Wherein x is the urea concentration and y is the current.
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Application publication date: 20210413 |