CN109613084B - High-sensitivity detection H of nano gold-protoporphyrin zinc (II)2O2Construction and application of electrochemical sensor - Google Patents

High-sensitivity detection H of nano gold-protoporphyrin zinc (II)2O2Construction and application of electrochemical sensor Download PDF

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CN109613084B
CN109613084B CN201811511431.9A CN201811511431A CN109613084B CN 109613084 B CN109613084 B CN 109613084B CN 201811511431 A CN201811511431 A CN 201811511431A CN 109613084 B CN109613084 B CN 109613084B
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zinc
protoporphyrin
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杨怀霞
刘艳菊
李晓飞
李金鸽
李曼曼
郭文峰
赵可欣
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Henan University of Traditional Chinese Medicine HUTCM
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Abstract

The invention discloses a high-sensitivity detection method for H by using nano gold-protoporphyrin zinc (II)2O2The construction and application of electrochemical sensor, in particular to the synthesis of nano gold-protoporphyrin zinc (II) (PEI-AuNPs-ZnPP) and H in human serum2O2Detection of (3). The method is synthesized by utilizing the characteristic that nanogold and protoporphyrin zinc can be connected through amido bonds, the nanogold and protoporphyrin zinc are modified on the surface of a glassy carbon electrode, and H is detected by adopting an electrochemical method2O2The concentration and the result show that under the optimal condition, the H is within a certain range2O2The concentration of (d) shows positive correlation with the current signal, the linear equation is I (μ a) ═ 0.20412c (pmol/L) +1.93557, the correlation coefficient is 0.99534, and the detection limit is 8.540 × 10‑13And M. The stability of the sensor is better expressed in that the current signal of the sensor is 96.8 percent of the initial value after 3 days, and simultaneously the sensor measures H in the serum of the actual sample human2O2Also shows higher recovery rate (97.57-101.43%), and can be used for detecting H2O2The concentration of (2) lays a foundation for later application in the medical field for detecting the concentration of glucose.

Description

High-sensitivity detection H of nano gold-protoporphyrin zinc (II)2O2Construction and application of electrochemical sensor
Technical Field
The invention belongs to the field of construction and application of electrochemical sensors, and particularly relates to synthesis of nanogold-protoporphyrin zinc (II) (PEI-AuNPs-ZnPP) and H in human serum2O2Belongs to the technical field of biological analysis.
Background
Hydrogen peroxide (H)2O2) As an important biomarker, the method is widely applied to the fields of food production, environmental monitoring, drug synthesis, clinical detection and the like, hydrogen peroxide is a toxic byproduct generated in the metabolic process of a human body and can cause huge damage to the body, and researches show that H2O2Overdose can cause alzheimer's disease, parkinson's disease and other central nervous system disorders; thus H2O2Sensitive detection of content is of great importance in the physiological, pathological and environmental fields. At present, there are many methods for measuring the hydrogen peroxide content, such as colorimetry, spectrophotometry, chemiluminescence, and electrochemical methods. However, most of these methods have the disadvantages of high detection cost, complicated operation, long analysis time, low detection limit, and the like. Therefore, H with high sensitivity and low detection limit is designed and developed2O2The detection method is necessary.
The nanogold (PEI-AuNPs) has the advantages of good biocompatibility, large specific surface area, large surface active center, high catalytic efficiency, strong adsorption capacity, high surface activity and the like, and is applied to the field of electrochemical sensors for standby application. At present, the preparation of nano gold is mostly based on HAuCl4And (3) reduction reaction of (2). Research shows that the Polyethyleneimine (PEI) is used as a reducing agent and a stabilizing agent to prepare the nano gold particles, the synthetic method is simple, and the green chemical concept is met.
The electrochemical method is favored by researchers due to the advantages of sensitive detection, high accuracy, low cost, simple operation and the like, and meanwhile, the electrochemical sensor becomes an active research field of electroanalytical chemistry and is widely applied to the fields of food and drug analysis, environmental monitoring, life science and the like. The nano gold-protoporphyrin zinc (II) electrochemical transmission in the inventionThe sensor has the advantages of high sensitivity, good selectivity, low detection limit and the like, and can be used for low-concentration H2O2And (6) detecting.
Disclosure of Invention
Aiming at the problems existing at present, the invention aims to provide the nanogold-protoporphyrin zinc (II) high-sensitivity H detection2O2Construction and application of electrochemical sensor.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a nano gold-protoporphyrin zinc (II) bionic enzyme is prepared by the following steps:
(1) ZnPP is dissolved in DMF to prepare ZnPP solution with the concentration of 0.1 mmol/L;
(2) mixing the ZnPP solution and the nano gold solution according to the volume ratio of 1.2:1, and stirring for 1 h;
(3) after stirring, centrifuging;
(4) discarding the supernatant, washing the precipitate, and centrifuging;
(5) discarding the supernatant, vacuum drying the precipitate, adding methanol after drying, and storing at 4 deg.C for use.
The preparation method of the nano gold solution comprises the following steps: weighing 2.06g of PEI and 340 mu L of HAuCl with the mass fraction of 2%4Mixing the solutions, adding 1960mL of ultrapure water, intermittently heating to 80 deg.C while stirring, observing the color change of the solution, stopping heating when the solution turns to light ruby red, and continuously stirring to room temperature.
The intermittent heating is as follows: heating from room temperature to 80 deg.C, and keeping the temperature for 1-2min every time the temperature is raised to 5 deg.C.
The temperature of centrifugation in the step (3) is 4 ℃, the rotating speed is 14000rpm, and the time is 75-85 min.
The temperature of centrifugation in the step (4) is 4 ℃, the rotating speed is 14000rpm, and the time is 40-50 min.
Preparation of H by using nano gold-protoporphyrin zinc (II) biomimetic enzyme2O2Use in an electrochemical sensor.
Preparation of H based on nanogold-protoporphyrin zinc (II) biomimetic enzyme2O2The method for preparing the electrochemical sensor comprises the following steps:
(1) cleaning of the electrode:
polishing the glassy carbon electrode to a mirror surface, then putting the polished glassy carbon electrode into an ultrasonic cleaner, respectively cleaning with ultrapure water, absolute ethyl alcohol and ultrapure water, and blow-drying with nitrogen for later use;
(2) modification of the electrode:
dripping the nano gold-protoporphyrin zinc (II) biomimetic enzyme on the surface of a glassy carbon electrode, and drying in vacuum at room temperature.
Vacuum drying for 30-50 min.
H detection of electrochemical sensor prepared by method in human serum2O2The use of (1).
The preparation method of protoporphyrin zinc refers to a synthesis method of a porphyrin zinc complex (ZnTpMoPP) in polyacrylic acid modified chitosan nanosphere zinc-porphyrin conjugate and performance research (Xuanji38881, university of northwest, 2010).
The preparation method and the detection principle of the electrochemical sensor are schematically shown in figure 1.
The invention has the beneficial effects that:
1. the PEI has better reducibility and stability to the gold particles, so that the prepared nano gold has small particle size and uniform distribution, the nano gold particles can be prevented from being precipitated, and the method is simple and convenient to operate and has mild reaction conditions.
2. The nanogold has good chemical stability, good conductivity, good biocompatibility and large specific surface area, and can promote H2O2And the transfer of electrons between the electrodes expands current signals, thereby improving the catalytic efficiency of the biomimetic enzyme.
3. The nanogold-protoporphyrin zinc (II) and hydrogen peroxide are subjected to oxidation-reduction reaction to generate an electric signal, and the sensor has the advantages of low detection limit, high sensitivity, high stability, good repeatability and the like.
4. The high-sensitivity nanogold-protoporphyrin zinc (II) is synthesized by utilizing the characteristic that nanogold and protoporphyrin zinc can be connected through amido bond, the nanogold and protoporphyrin zinc are modified on the surface of a glassy carbon electrode, and H is detected by adopting an electrochemical method2O2The concentration and the result show that under the optimal condition, the H is within a certain range2O2The concentration of (d) shows positive correlation with the current signal, the linear equation is I (μ a) ═ 0.20412c (pmol/L) +1.93557, the correlation coefficient is 0.99534, and the detection limit is 8.540 × 10-13And M. The stability of the sensor is better expressed in that the current signal of the sensor is 96.8 percent of the initial value after 3 days, and simultaneously the sensor measures H in the serum of the actual sample human2O2Also shows higher recovery rate (97.57-101.43%), and can be used for detecting H2O2The concentration of (2) lays a foundation for later application in the medical field for detecting the concentration of glucose.
Drawings
FIG. 1 is a schematic diagram of the preparation method and detection principle of the electrochemical sensor of the present invention.
FIG. 2(a) is a TEM image of PEI-AuNPs.
FIG. 2(b) is a UV-visible absorption spectrum of PEI-AuNPs.
FIG. 3(a) is a TEM image of PEI-AuNPs-ZnPP biomimetic enzyme.
FIG. 3(b) is a Fourier infrared spectrum of PEI-AuNPs-ZnPP biomimetic enzyme.
FIG. 4 shows the effect of different modified electrodes on the current signal.
FIG. 5 is a contact angle for different modified electrodes; in the figure, (a): bare electrode (b): PEI-AuNPs (c): ZnPP (d): PEI-AuNPs-ZnPP.
Fig. 6 shows the effect of different scan rates on the current signal and the relationship of the peak current to the scan rate (V/s) (inset).
FIG. 7 is a graph of peak current versus pH.
FIG. 8 shows the relationship between peak current and modification amount.
FIG. 9 is a graph of peak current versus trim time.
FIG. 10(a) shows a difference H2O2The effect of concentration on the current signal; in the figure H2O2The concentration of (A) is 0.30, 0.35, 0.40, 0.45, 0.50, 0.80, 1.00, 1.30, 1.50, 1.80, 2.00, 2.10, 2.60, 3.00, 3.50, 4.00, 4.50, 5.50, 5.80, 5.90, 6.00pmol/L in sequence from top to bottom.
FIG. 10(b) shows the peak current and H2O2Concentration C (pmol/L);in the figure H2O2The concentration of (A) is 0.50, 0.80, 1.00, 1.30, 1.50, 1.80, 2.10, 2.60 and 3.00pmol/L in sequence from left to right.
FIG. 11 shows the change in peak current of the sensor before and after the addition of the interfering substance (a 1: H)2O2，a2：H2O2+ dopamine, b 1: h2O2，b2：H2O2+ uric acid, c 1: h2O2，c2：H2O2+ glucose).
Detailed Description
The following examples further illustrate the embodiments of the present invention in detail.
Example 1: synthesizing nano gold-protoporphyrin zinc (II) (PEI-AuNPs-ZnPP) biomimetic enzyme:
(1) 2.06g of Polyethyleneimine (PEI) was weighed out together with 340. mu.L of 2% by mass chloroauric acid (HAuCl)4) Mixing the solutions, adding 1960mL of ultrapure water, intermittently heating to 80 ℃ while stirring, keeping the temperature, observing the color change of the solution, stopping heating when the solution becomes light ruby red, continuously stirring to room temperature to obtain a solution of the nanogold (PEI-AuNPs), and storing at 4 ℃ in a dark place for later use;
(2) dissolving protoporphyrin zinc (II) (ZnPP) in N, N-Dimethylformamide (DMF) to prepare a ZnPP solution with the concentration of 0.1 mmol/L;
(3) mixing 4.8mL of ZnPP solution with 4mL of nano-gold solution, and stirring the mixed solution on a magnetic stirrer for 1 h;
(4) after stirring, transferring the mixed solution into a centrifuge tube, and centrifuging for 80min at 14000rpm at 4 ℃;
(5) after centrifugation, removing supernatant, adding 1.0mL of ultrapure water into a centrifuge tube to wash precipitates, and centrifuging for 40min at 14000rpm at 4 ℃;
(6) after centrifugation, the supernatant was discarded, and the precipitate was vacuum dried at room temperature, and after drying, 150. mu.L of methanol was added to the centrifuge tube and stored at 4 ℃ for further use.
The intermittent heating method comprises the following steps: the temperature is raised from room temperature to 80 ℃, and the temperature is kept for 2min every time the temperature is raised to 5 ℃.
Structural characterization of the nanogold:
(1) morphology and particle size characterization
The particle size of PEI-AuNPs was measured by a nanometer particle size Zeta potentiometer (Malvern, UK), and the morphological distribution of PEI-AuNPs was observed by a Japanese Hitachi Transmission Electron Microscope (TEM), as shown in FIG. 2(a), the average diameter of nanogold was 10nm, and the nanogold was spherical and uniformly distributed.
(2) Characterization of UV-visible absorption Spectroscopy
The UV-3600Plus (SHIMADZU, Japan) ultraviolet-visible absorption spectrometer is adopted to characterize the synthesized PEI-AuNPs, and the result is shown in figure 2(b), and the PEI-AuNPs have a characteristic absorption peak at 525nm, so that the PEI-AuNPs can be judged to be successfully prepared.
Structural characterization of the nanogold-protoporphyrin zinc (II):
(1) transmission Electron Microscope (TEM) analysis
The PEI-AuNPs-ZnPP was analyzed by a Transmission Electron Microscope (TEM) of Hitachi Japan, and the average diameter of the PEI-AuNPs-ZnPP was 10nm as shown in FIG. 3 (a).
(2) Fourier transform infrared spectroscopy
Performing infrared spectrum test (liquid smear method, scanning at room temperature, and test range of 1000-4000 cm) on the obtained PEI-AuNPs-ZnPP compound by NICOLET iS50 Fourier transform infrared spectrometer manufactured by American Siemens fly (Thermo SCIENTIFIC)-1) The results are shown in FIG. 3(b), 1454.26cm-1Is due to N-H in-plane vibration at 1658.10 cm-1The peak is due to stretching vibration of C ═ O, which shows that amide bond connection between the nano gold and the zinc (II) protoporphyrin is successful, so that the nano gold-zinc (II) protoporphyrin can be judged to be successfully prepared.
Example 2: preparation of electrochemical sensor
(1) Cleaning of the electrode (GCE):
on a polishing plate, polishing a glassy carbon electrode to a mirror surface by using alumina polishing powder with the particle size of 0.3 mu m, and then inserting the glassy carbon electrode into a 5mmol/L potassium ferricyanide solution to carry out a linear cyclic voltammetry scanning test (the scanning range is-0.2V-0.6V, the scanning rate is 0.05V/s, the sampling interval is 1mV, the static time is 5s, the cathode current is positive, and the electrode is considered to be qualified when the redox peak potential difference of the glassy carbon electrode is less than 80mV, and if the electrode is not qualified, the operation is repeated). And (3) placing the glassy carbon electrode qualified in the test into an ultrasonic cleaner, sequentially cleaning with ultrapure water, absolute ethyl alcohol and ultrapure water for 1min, and finally drying with nitrogen.
(2) Modification of the electrode:
dripping 4-6 mu L of nano gold-protoporphyrin zinc (II) on the surface of the glassy carbon electrode, and drying in vacuum for 30-50min at room temperature.
Example 3: electrochemical sensor feasibility detection
Respectively modifying 5 mu L of nanogold (PEI-AuNPs), zinc (II) protoporphyrin (ZnPP) and nanogold-zinc (II) protoporphyrin (PEI-AuNPs-ZnPP) on the surface of an electrode (the method is the same as that of example 2), carrying out vacuum drying on the electrode and a naked electrode for 30min, and adding 10 mu L of 0.5mol/L H into 20mL of PBS buffer solution with the pH value of 7.000.2 mol/L2O2And differential pulse voltammetry is used for measurement (scanning range: 0V to-1V), the result is shown in figure 4, and the current signal of the nano gold-zinc protoporphyrin (II) is strongest, so that the method is proved to be feasible.
Example 4: electrochemical sensor contact angle measurement
Respectively modifying 5 mu L of nanogold (PEI-AuNPs), zinc (II) protoporphyrin (ZnPP) and nanogold-zinc (II) protoporphyrin (PEI-AuNPs-ZnPP) on the surface of a glassy carbon electrode (the method is the same as that in example 2), carrying out vacuum drying on the glassy carbon electrode and a bare electrode for 30min, and measuring the contact angle of the glassy carbon electrode and the bare carbon electrode by using a video optical contact angle measuring instrument, wherein the contact angles are different when the surface of the electrode is modified by different materials as shown in figure 5.
Example 5: detection of sensor reaction type
Using the sensor of example 2, 10. mu.L of 0.1mol/L H was added to a PBS buffer solution at pH 7.500.2 mol/L20mL2O2The sweep rates (0.01V/s, 0.02V/s, 0.05V/s, 0.10V/s, 0.15V/s, 0.20V/s, 0.25V/s and 0.30V/s) are selected to be measured by linear cyclic voltammetry (the sweep range is between-0.9V and 1.2V), the result is shown in figure 6, the linear equation I (mu A) is between 0.7924 and 22.03598V (V/s), the correlation coefficient is 0.99109, and the current signal is positively correlated with the sweep rate, which indicates the hydrogen peroxide cleaning performance of the sensor of the inventionThe detection belongs to a surface control process.
Example 6: optimization of detection conditions
(1) Optimization of pH
Using the biosensor of example 2, 10. mu.L of 0.5mol/L H was added to 0.2mol/L of 20mL of PBS buffer (pH 6.00, 6.50, 7.00, 7.50, 7.75, 8.50)2O2As a result of measurement by differential pulse voltammetry (scanning range: 0V to-1V), pH is preferably 7.5 as shown in FIG. 7.
(2) Optimization of modification amounts
Modifying 2 μ L, 2.5 μ L, 3 μ L, 4 μ L, 5 μ L, 6 μ L of nano gold-protoporphyrin zinc (II) on a dry and clean glassy carbon electrode, and vacuum drying for 30 min. To 20mL of 0.2mol/L PBS buffer solution (pH 7.5), 10. mu.L of 0.5mol/L H was added2O2As a result of measurement by differential pulse voltammetry (scanning range: 0V to-1V), the modification amount is preferably 4. mu.L as shown in FIG. 8.
(3) Optimization of modification time
mu.L of nanogold-protoporphyrin zinc (II) was modified on the electrode surface, vacuum dried (modification time) (10, 20, 30, 40, 50, 60, 70) respectively, and 10. mu.L of 0.5mol/L H was added to 20mL of 0.2mol/L PBS buffer solution (pH 7.5)2O2As a result of measurement by differential pulse voltammetry (scanning range: 0V to-1V), the modification time is preferably 40min as shown in FIG. 9.
From this, the optimal optimization conditions are: the pH of the PBS buffer solution was 7.5, the modification amount of nanogold-protoporphyrin zinc (ii) was 4 μ L, and the modification time was 40 min.
Example 7: linear range detection for electrochemical sensors
According to the optimal experimental conditions obtained by optimizing the conditions of the embodiment 6, 4 mu L of nano gold-zinc protoporphyrin (II) is modified on a dry and clean glassy carbon electrode and is dried for 40min in vacuum. To 20mL of 0.2mol/L PBS buffer (pH 7.5) were added different amounts (60 μ L, 70 μ L, 80 μ L, 90 μ L, 100 μ L, 160 μ L, 220 μ L, 260 μ L, 320 μ L, 400 μ L, 500 μ L, 600 μ L, 800 μ L, 900 μ L, 1100 μ L, 1160 μ L, 1180 μ L, 1200 μ L) of 1 × 10 μ L-10mol/L hydrogen peroxide, then using the differenceThe pulsed voltammetry was shown to measure the current intensity of the electrode versus different concentrations of hydrogen peroxide (fig. 10). The relationship between the current intensity and the hydrogen peroxide concentration is researched, a linear range (0.5 pmol/L-3.0 pmol/L) and a linear regression equation I (mu A) of 0.20412c (pmol/L) +1.93557 are obtained, the correlation coefficient is 0.99534, and the detection limit is 8.540 × 10-13M。
Example 8: selective detection of electrochemical sensors
According to the optimal experimental conditions obtained by optimizing the conditions of the embodiment 6, 4 mu L of nano gold-zinc protoporphyrin (II) is modified on a dry and clean glassy carbon electrode and is dried for 40min in vacuum. To 20mL of 0.2mol/L PBS buffer solution (pH 7.5) was added 10. mu.L of 1X 10-10mol/L of H2O2Then 10. mu.L of 5X 10 were added separately-9mol/L Dopamine (DA), 10. mu.L 5X 10-9mol/L Glucose (GC), 10. mu.L 5X 10-9mol/L Uric Acid (UA), and then measuring the current intensity (scanning range: 0V to-1V) of the electrode to hydrogen peroxide solution with different concentrations by using differential pulse voltammetry (FIG. 11). The result shows that the addition of Dopamine (DA), Glucose (GC) and Uric Acid (UA) can detect H for PEI-AuNPs-ZnPP2O2Basically has no influence, so the PEI-AuNPs-ZnPP sensor pair H2O2The detection has good selectivity.
Example 9: stability and repeatability detection of electrochemical sensors
According to the optimal experimental conditions obtained by optimizing the conditions of the embodiment 6, 4 mu L of nano gold-protoporphyrin zinc (II) is modified on the surface of the glassy carbon electrode, and vacuum drying is carried out for 40 min. To 20mL of 0.2mol/L PBS buffer solution (pH 7.5), 10. mu.L of 0.5mol/L H was added2O2And differential pulse voltammetry is used for measurement (the scanning range is 0V to-1V), and the result shows that the detection signal of the sensor to hydrogen peroxide with the same concentration reaches 96.8 percent of the initial detection after the sensor is placed for three days, which indicates that the sensor has better stability. According to the error bars of each group of data, the electrochemical sensor has good repeatability.
Example 10: application of electrochemical sensor in human serum
Based on the optimal experimental conditions obtained by optimizing the conditions of example 6, 4Mu L of nano gold-protoporphyrin zinc (II) is decorated on a dry and clean glassy carbon electrode and is dried for 40min in vacuum. To 20mL of 0.2mol/LPBS buffer solution (pH 7.5) containing different volume percentages of serum (1%, 5%, 10%) was added 100 μ L of different concentrations (2 × 10)-10mol/L、4×10-10 mol/L、5×10-10mol/L) of H2O2The results are shown in Table 1, where the recovery rate is 97.57-101.43% compared to the current signal of the same concentration in the linear phase.
TABLE 1 detection of human serum samples with PEI-AuNPs-ZnPP composite (n. 3)
Figure BDA0001900887930000071

Claims (8)

1. A nano gold-protoporphyrin zinc (II) bionic enzyme is characterized in that the preparation method comprises the following steps:
(1) ZnPP is dissolved in DMF to prepare ZnPP solution with the concentration of 0.1 mmol/L;
(2) mixing the ZnPP solution and the nano gold solution according to the volume ratio of 1.2:1, and stirring for 1 h;
(3) after stirring, centrifuging;
(4) discarding the supernatant, washing the precipitate, and centrifuging;
(5) discarding the supernatant, vacuum drying the precipitate, adding methanol after drying, and storing at 4 deg.C for use;
the preparation method of the nano gold solution comprises the following steps: weighing 2.06g of PEI and 340 mu L of HAuCl with the mass fraction of 2%4Mixing the solutions, adding 1960mL of ultrapure water, intermittently heating to 80 deg.C while stirring, observing the color change of the solution, stopping heating when the solution turns to light ruby red, and continuously stirring to room temperature.
2. The nanogold-zinc protoporphyrin (II) biomimetic enzyme according to claim 1, wherein the intermittent temperature rise is as follows: heating from room temperature to 80 deg.C, and keeping the temperature for 1-2min every time the temperature is raised to 5 deg.C.
3. The nanogold-protoporphyrin zinc (II) biomimetic enzyme according to claim 1, wherein the temperature of centrifugation in step (3) is 4 ℃, the rotation speed is 14000rpm, and the time is 75-85 min.
4. The nanogold-protoporphyrin zinc (II) biomimetic enzyme according to claim 1, wherein the temperature of centrifugation in step (4) is 4 ℃, the rotation speed is 14000rpm, and the time is 40-50 min.
5. Preparation of H by using nano gold-protoporphyrin zinc (II) biomimetic enzyme as claimed in claim 12O2Use in an electrochemical sensor.
6. Preparation of H based on nanogold-protoporphyrin zinc (II) biomimetic enzyme of claim 12O2The method for preparing the electrochemical sensor is characterized by comprising the following steps:
(1) cleaning of the electrode:
polishing the glassy carbon electrode to a mirror surface, then putting the polished glassy carbon electrode into an ultrasonic cleaner, respectively cleaning with ultrapure water, absolute ethyl alcohol and ultrapure water, and blow-drying with nitrogen for later use;
(2) modification of the electrode:
dripping the nano gold-protoporphyrin zinc (II) biomimetic enzyme on the surface of a glassy carbon electrode, and drying in vacuum at room temperature.
7. Preparation of H on the basis of nanogold-protoporphyrin zinc (II) biomimetic enzyme according to claim 62O2The method of the electrochemical sensor is characterized in that the vacuum drying time is 30-50 min.
8. An electrochemical sensor prepared according to the method of claim 6 for detecting H in human serum2O2The use of (1).
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