CN114441492A - Method for detecting hydrogen peroxide by enzyme-labeling instrument - Google Patents
Method for detecting hydrogen peroxide by enzyme-labeling instrument Download PDFInfo
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Abstract
The invention relates to the technical field of detection, in particular to a method for detecting hydrogen peroxide by an enzyme-labeling instrument. Which comprises the following steps: dividing the mixed solution of the nitrogen-doped carbon quantum dot solution, the ethanol solution of tetramethylbenzidine and the acetate buffer solution with pH value of 3.2 into more than 3 parts, respectively adding hydrogen peroxide solutions with different concentrations, reacting at 35 ℃ for 15-25 min, transferring to a pore plate, placing into an enzyme labeling instrument for detection, and making a standard curve; adding a hydrogen peroxide solution to be detected into the mixed solution with the same concentration, reacting for 15-25 min at 35 ℃, transferring into a pore plate, placing into an enzyme-labeling instrument for detection, and determining the concentration of the hydrogen peroxide solution to be detected. The method for detecting the hydrogen peroxide by the enzyme-labeling instrument has the advantages of low cost, rapidness, simplicity, convenience, sensitivity and specificity because the nitrogen-doped quantum dots (N-CDs) are used as the hydrogen peroxide mimic enzyme and the hydrogen peroxide is quantitatively detected by the enzyme-labeling instrument.
Description
Technical Field
The invention relates to the technical field of detection, in particular to a method for detecting hydrogen peroxide by an enzyme-labeling instrument.
Background
Hydrogen peroxide (H)2O2) Commonly known as hydrogen peroxide, a strong oxidant with wide application field. In environmental remediation, H2O2Are often used to treat industrial and domestic wastewater. However, the excessive consumption of the hydrogen peroxide can cause environmental pollution and influence the normal growth and metabolic activities of microorganisms in the water body, and once the hydrogen peroxide in the water body enters the human body, the health of the human body can be damaged. In the human body, H2O2Exists in the form of active oxygen, participates in various physiological metabolic processes of human body, and has high concentration of H2O2Can disturb the normal metabolism of the human body and even cause cell damage. Thus, the detection of hydrogen peroxide is of great practical significance. The current methods for detecting hydrogen peroxide mainly include titration, electrochemical methods, photometric methods, fluorescence/chemiluminescence methods, refractive index methods and microwave methods. The chromogenic method generally uses Tetramethylbenzidine (TMB) as a color-developing agent to measure horseradish peroxidase and the like, and is simple to operate and has high sensitivity and a low detection limit. However, biological enzymes are easy to inactivate and high in cost, and the search for artificial enzymes becomes a current research hotspot.
Carbon Quantum Dots (CQDs) are a novel carbon nano fluorescent material, have the characteristics of low manufacturing cost, good biological stability, good photochemical performance and low cytotoxicity, are generally below 10nm in size, contain rich oxygen-containing functional groups on the surface, have good hydrophilic performance and strong controllability, and can be used for detecting related substances. At present, the synthesis methods of the carbon quantum dots comprise hydrothermal synthesis methods, electrochemistry methods, chemical oxidation methods and the like, and the microwave method is a time-saving, labor-saving, rapid and simple preparation method and is widely concerned by researchers. For example, Wangshiqi and the like adopt a microwave method to prepare N-CDs for exploring the rapid preparation of nitrogen-doped carbon dots and are used as probes for detecting iron ions. Limanxiu and the like adopt a microwave method to prepare a carbon quantum dot and chitosan which are compounded for detecting quercetin.
The patent with the publication number of CN202110468257.X discloses a nitrogen-doped graphene quantum dot, and preparation and application thereof in hydrogen peroxide detection, wherein the nitrogen-doped graphene quantum dot is used as an anode electrochemical luminescence probe for detecting hydrogen peroxide. The carbon source compound is 1, 3-dinitronaphthalene, and the dopant is luminol.
Disclosure of Invention
Technical problem to be solved
In view of the above disadvantages and shortcomings of the prior art, the invention provides a method for detecting hydrogen peroxide by using nitrogen-doped carbon quantum dots as an enzyme reader of hydrogen peroxide mimic enzyme, which is a detection method with simple operation, low cost and high speed.
(II) technical scheme
In order to achieve the purpose, the invention adopts the main technical scheme that:
in a first aspect, an embodiment of the present invention provides a method for detecting hydrogen peroxide by a microplate reader, which includes the following steps:
s1 standard curve preparation: dividing the mixed solution of the nitrogen-doped carbon quantum dot solution, the ethanol solution of tetramethylbenzidine and the acetate buffer solution with pH value of 3.2 into more than 3 parts, respectively adding hydrogen peroxide solutions with different concentrations, reacting at 35 ℃ for 15-25 min, transferring to a pore plate, placing into an enzyme labeling instrument for detection, and making a standard curve;
and S2, adding the hydrogen peroxide solution to be detected into the mixed solution with the same concentration as that in the step S1, reacting at 35 ℃ for 15-25 min, transferring into a pore plate, placing into an enzyme labeling instrument, detecting, and determining the concentration of the hydrogen peroxide solution to be detected.
In the optional scheme of the method for detecting hydrogen peroxide by the microplate reader, the concentration of citric acid in the mixed solution is 0.04-0.06 g/ml, and the concentration of urea is 0.02-0.04 g/ml.
In the optional scheme of the method for detecting hydrogen peroxide by the microplate reader, the preparation method of the nitrogen-doped carbon quantum dots comprises the following steps: and (2) carrying out microwave treatment on the mixed solution of citric acid and urea in microwave for 2.5-4 min, adding ultrapure water for dissolving, centrifuging, taking supernatant, filtering with a 0.22-micron filter membrane, and drying the dialyzed solution in a 1000Kd dialysis bag to obtain the nitrogen-doped carbon quantum dot.
(III) advantageous effects
The invention has the beneficial effects that: according to the method for detecting the hydrogen peroxide by the enzyme-labeling instrument, nitrogen-doped quantum dots (N-CDs) are used as the hydrogen peroxide mimic enzyme, Tetramethylbenzidine (TMB) is used as a color developing agent, and quantitative hydrogen peroxide detection is carried out by the enzyme-labeling instrument, so that the method is low in cost, rapid, simple and convenient, has sensitivity and specificity, and can be used for detecting the hydrogen peroxide in a water body.
According to the invention, the nitrogen-doped carbon quantum dots prepared from the mixed solution of citric acid and urea are used for detecting hydrogen peroxide, and compared with the nitrogen-doped graphene quantum dots disclosed in the patent of CN202110468257.X, the nitrogen-doped carbon quantum dots have significantly better stability.
Drawings
FIG. 1 is an absorption diagram of ultraviolet spectra of N-CDs obtained at four different microwave times;
FIG. 2 is a photograph of an aqueous solution of carbon dots under daily light of N-CDs and a photograph of a fluorescent image under irradiation of a 365nm ultraviolet lamp, which are obtained at four different microwave times;
FIG. 3 is an absorption diagram of UV spectra of N-CDs obtained at four different microwave times;
FIG. 4 is a graph of N-CDs aqueous solution scanned in Fourier infrared spectroscopy at four different microwave times;
FIG. 5 is an XRD representation of the N-CDs powder obtained by microwave 3.5 min;
FIG. 6 is an XRD representation of the N-CDs powder obtained by microwave for 4 min;
FIG. 7 is an XPS characterization of N-CDs powder obtained by microwave 4 min;
FIG. 8 is a C1s high resolution spectrum of N-CDs powder obtained by microwave for 4 min;
FIG. 9 is a high resolution N1s spectrum of N-CDs powder obtained by microwave for 4 min;
FIG. 10 is a high resolution O1s spectrum of N-CDs powder obtained by microwave 4 min;
FIG. 11 shows the catalytic effect of N-CDs on hydrogen peroxide obtained by microwave for 3.5 min;
FIG. 12 is a graph showing the catalytic effect of N-CDs on hydrogen peroxide at different microwave times;
FIG. 13 shows the results of hydrogen peroxide measurements at different N-CDs concentrations;
FIG. 14 shows the results of hydrogen peroxide measurements at different TMB concentrations;
FIG. 15 shows the results of hydrogen peroxide measurements at different reaction pH;
FIG. 16 shows the results of hydrogen peroxide measurements at different reaction temperatures;
FIG. 17 shows the results of hydrogen peroxide measurements at different reaction times;
FIG. 18 is a graph showing the color change of hydrogen peroxide concentration measured by UV spectrophotometry;
FIG. 19 is a standard curve of 0.5-200 μmol/L hydrogen peroxide concentration measured by UV spectrophotometry;
FIG. 20 is a standard curve of 0.5-2.5 μmol/L hydrogen peroxide concentration measured by UV spectrophotometry;
FIG. 21 is a standard curve of hydrogen peroxide concentration of 100-200 μmol/L measured by UV spectrophotometry; FIG. 22 is a diagram showing that 0.5 to 200. mu. mol/L reaction is carried out for 20min by an enzyme-labeling instrument for H determination2O2And the OD value;
FIG. 23 is a diagram showing that H is measured by a 20min enzyme-labeling instrument through 0.5-2.5 mu mol/L reaction2O2And the OD value;
FIG. 24 shows the measurement of H by a 100-200. mu. mol/L reaction 20min enzyme-labeling instrument2O2And the OD value;
FIG. 25 is a diagram showing that H is measured by a microplate reader with a reaction time of 0.5 to 200. mu. mol/L for 15min2O2And the OD value;
FIG. 26 is a diagram showing that H is measured by a 0.5-5 mu mol/L reaction 15min enzyme-labeling instrument2O2And the OD value;
FIG. 27 shows the measurement of H by a 25min microplate reader with 0.5-200. mu. mol/L reaction2O2And the OD value;
FIG. 28 is a diagram showing the measurement of H by a 25min enzyme-linked immunosorbent assay (ELIASA) for 50-200. mu. mol/L reaction2O2And a linear plot between OD values.
Detailed Description
For a better understanding of the present invention, reference will now be made in detail to the present invention by way of specific embodiments thereof.
The invention provides a novel, rapid, low-cost, sensitive and specific method for detecting hydrogen peroxide in a water body, which takes citric acid and urea as raw materials, synthesizes nitrogen-doped quantum dots (N-CDs) by a microwave method to be used as hydrogen peroxide mimic enzyme, takes Tetramethylbenzidine (TMB) as a color developing agent, and carries out quantification by an enzyme-labeling instrument.
The nitrogen-doped carbon quantum dots are preferably prepared by taking citric acid and urea as raw materials and adopting a simple microwave method.
In order to better understand the above technical solutions, exemplary embodiments of the present invention will be described in more detail below. While the following shows exemplary embodiments of the invention, it should be understood that the invention may be embodied in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
Example 1
Preparation of nitrogen-doped carbon quantum dots
Dissolving 2.5g of citric acid and 1.5g of urea in 50mL of ultrapure water, magnetically stirring at normal temperature for 10min to form a uniform solution, placing 10mL of 4 parts of mixed solution in a microwave oven respectively, performing microwave treatment for 2.5min, 3min, 3.5min and 4min to obtain a brownish black substance, naturally cooling to room temperature, adding an appropriate amount of ultrapure water for dissolving, centrifuging for 10min (10000r/min), filtering a supernatant with a 0.22 mu m filter membrane, and adding ultrapure water for diluting to 100 mL. Putting the solution into a 1000Kd dialysis bag, dialyzing for 24h, changing water every 3h, and washing and purifying to obtain a pure N-CDs solution; and (3) placing the N-CDs solution in a freeze dryer for freeze drying for 24h to obtain brown nitrogen-doped carbon quantum dot powder.
The N-CDs carbon quantum dots obtained in this example were analyzed as follows:
1. fluorescence spectroscopy of carbon quantum dots
FIG. 1 is a graph of the absorption of ultraviolet spectra of N-CDs at four different microwave times, and the fluorescence intensity at emission wavelengths from 350nm to 700nm was measured at a fixed excitation wavelength of 360 nm. N-CDs with the microwave time of 2.5min has an absorption broad peak at 450nm, and the fluorescence intensity of the carbon quantum dot is obviously stronger than that of carbon dots obtained in other microwave times. The absorption peak of N-CDs with microwave time of 3min is located at 500nm, and the absorption peaks of N-CDs with microwave time of 3.5min and 4min are located near 530 nm. The maximum absorption peak is red-shifted with increasing microwave time. This may be related to the particle size distribution of the carbon quantum dots, or may be caused by surface defects of the carbon quantum dots. The fluorescence absorption intensity of the carbon quantum dots decreases with increasing microwave time, because the fluorescence intensity decreases as the carbon dot particle size increases with increasing reaction time, due to the oversize carbon dots.
FIG. 2 is a photograph of an aqueous solution of carbon dots under a daily light and a photograph of a fluorescent image under the irradiation of a 365nm ultraviolet lamp, wherein the carbon quantum dot solutions with different microwave times have different fluorescence intensities. Meanwhile, the carbon quantum dot solutions are stored in a sealed and dark place, and the fluorescence intensity is tested again after 1 month, so that the fluorescence intensity is almost unchanged, and the long-term stability and good light stability of the fluorescence intensity of the carbon quantum dots can be seen.
2. Ultraviolet absorption spectroscopy analysis of carbon quantum dots
It can be seen from the figure that the absorption peak becomes wider after microwave heating for 3min at around 230nm, while fig. 3 is an absorption diagram of the ultraviolet spectrum of N-CDs for four different microwave heating times, and has a common absorption peak at 330nm, and the two absorption bands are formed by transitions of pi-pi and N-pi of C-C, C-O, C-N. As the microwave time is prolonged, the absorption peak at 330nm generates slight blue shift, and after microwave for 3.5min, a new peak position is gradually formed at 380nm, and the absorption in the visible light region is gradually increased.
3. Infrared absorption spectroscopy analysis of carbon quantum dots
FIG. 4 is a graph obtained by scanning four N-CDs aqueous solutions with different microwave times in a Fourier infrared spectrum, and the N-CDs peak positions at different microwave times are approximately the same by analyzing the surface structures of the four aqueous solutions. There is a concentrated absorption peak at 1456cm-1, which is due to the stretching vibration of C ═ C, constituting the basic unit of the carbon quantum dot. The peak at 1635cm-1 illustrates that the material may contain stretching vibrations of C ═ O. Broad absorption peaks appear at 3461cm-1 and 3243cm-1, which are O-H and N-H stretching vibrations.
XRD pattern analysis
Fig. 5 is an XRD representation of the N-CDs powder obtained by freeze drying, and as shown in fig. 5, N-CDs heated for 3.5min have a wide absorption peak at 25.2 degrees, which indicates that the structure is similar to the crystal face of graphene (002), and is an amorphous carbon structure.
As shown in fig. 6, N-CDs heated for 4min has a broad characteristic peak at 19.76 °, which also corresponds to the crystal plane of graphene (002). Therefore, the structures of the carbon quantum dots synthesized in the experiment are all graphite-like. The reason for generating the wide diffraction peak is that the prepared quantum dot has small grain diameter, low crystallinity and good dispersibility[8]. According to the Scherrer formula, the particle sizes of the synthesized N-CDs are all below 5nm and the sizes are extremely small.
XPS Spectroscopy
XPS characterization of N-CDs powder with microwave time of 4min showed that three characteristic peaks at 285.08, 400.08 and 532.08eV correspond to C1s, N1s and O1s respectively, as shown in FIG. 7, indicating that the N-CDs are composed of C, N, O element, further indicating that the carbon quantum dots are successfully doped with nitrogen element. Peak fitting was performed on the high resolution spectra of C1s, N1s, and O1 s.
As in fig. 8, three fitted peaks were obtained from the peak C1s, C — C/C ═ C (284.68eV), C-O/C-N (286.43eV), and C ═ O (288.08eV), respectively.
As shown in FIG. 9, two fitted peaks of C-N (339.63eV) and N-H (401.93eV) were isolated from N1s, indicating that the nitrogen in the carbon quantum dot is predominantly in the form of a C-N group.
As shown in fig. 10, the peak fitting of C1s resulted in two peaks, COOH/C ═ O — C (531.33eV) and C ═ O (532.53 eV). According to the characterization result of XPS, the synthesized carbon quantum dot surface has abundant carboxyl, hydroxyl and amino, so that N-CDs have good hydrophilicity.
Example 2
The detection of hydrogen peroxide comprises the following steps:
s1, adding 200 mu L of 150 mu g/L N-CDs solution, 600 mu L of 500 mu mol/LTMB ethanol solution, 200 mu L of a series of hydrogen peroxide solutions with different concentrations (0.1, 0.25, 0.5, 1, 1.5, 2, 2.5, 5, 10, 25, 50, 75, 100, 125, 150, 175 and 200 mu mol/L) and 1000 mu L of acetate buffer solution with pH3.2 into a centrifuge tube, placing the mixed solution into a pore plate after water bath for 20min at 35 ℃, placing the mixed solution into a microplate reader for detection, and establishing a standard curve; or measuring its absorbance at 652nm under an ultraviolet spectrophotometer;
s2 adding 200 mu L of 150 mu g/L N-CDs solution, 600 mu L of 500 mu mol/LTMB ethanol solution, 200 mu L of a series of hydrogen peroxide solutions with different concentrations (0.1, 0.25, 0.5, 1, 1.5, 2, 2.5, 5, 10, 25, 50, 75, 100, 125, 150, 175 and 200 mu mol/L) and 1000 mu L of pH3.2 acetate buffer solution into a centrifuge tube, reacting at 35 ℃ for 15-25 min, then putting 200 mu L into a 96-well plate, and placing the plate into a microplate reader for detection; or measuring its absorbance at 652nm under an ultraviolet spectrophotometer;
s3 taking 200uL of 150 mu g/L N-CDs solution, 600 mu L of 500 mu mol/L TMB ethanol solution, 200 mu L of hydrogen peroxide solution to be detected and 1000 mu L of acetate buffer solution with pH3.2, reacting for 15-25 min at 35 ℃, then putting 200 mu L into a 96-well plate, and placing into a microplate reader for detection.
The N-CDs obtained in example 1 were examined according to the method of this example to obtain the following results:
effect of catalytic Activity of N-CDs on color reaction
In order to prove the feasibility of the reaction, N-CDs with a microwave time of 3.5min were selected for the reaction in this example, and compared with the reaction system without N-CDs, the absorbance was measured after 10min of reaction at room temperature, and the results are shown in FIG. 11, where the absorbance value of the reaction system with N-CDs added was significantly higher than that of the reaction system without N-CDs, which proves that N-CDs have the catalytic activity of catalase. This is because N-CDs are rich in carbonyl groups or carboxyl groups on the surface, and promote adsorption of H2O2Electrocatalytic reduction of (c). That is, N-CDs have electron transfer capability, and lone electron pairs on TMB amino groups can be transferred to H through N-CDs2O2Thereby increasing H2O2The decomposition rate of (c);
2. respectively detecting the N-CDs aqueous solutions obtained in the example 1 at different microwave times by the method of the embodiment; as can be seen from FIG. 12, as the microwave time of the carbon spot increases, N-CDs vs. H2O2The catalytic activity of (2) is gradually increased, but the potential safety hazard is easily caused by too long heating time.
3. The N-CDs obtained by microwave 4min in example 1 were measured by the method of this example, and the concentration (50-150. mu.g/mL) of N-CDs, the concentration (50-500. mu. mol/L) of TMB, the pH (3.2-6.0), the temperature (25-45 ℃) and the time (10-45 min) were each measured as a single variable, while the other conditions were fixed. The results are shown in FIGS. 13-17:
as the concentration of N-CDs increases, the absorbance of the whole system at 652nm gradually increases, but if the concentration of N-CDs is too high, the reaction is too fast, so that a spectrophotometer cannot capture the absorbance. The absorbance gradually increased with the increase in TMB concentration, and reached the highest at a TMB concentration of 500. mu. mol/L. The pH is particularly important for the color reaction of the whole system, and the absorbance is maximum when the pH is 3.2 from pH3.2 to pH 6.0, and the absorbance of the whole system is continuously reduced along with the increase of the pH value. When the reaction temperature T is 35 ℃, the absorbance of the system is the maximum, and the absorbance of the system begins to decrease when the temperature is continuously increased. The absorbance increased with the increase of the reaction time, and the absorbance reached the maximum after the reaction for 30min, after which the absorbance began to decrease.
4. Ultraviolet spectrophotometry method for measuring hydrogen peroxide concentration
Preparation H2O2The standard solution of (4) has a concentration of 0.10, 0.25, 0.50, 1, 1.5, 2, 2.5, 5, 10, 25, 50, 75, 100, 125, 150, 175, 200. mu. mol/L, respectively. Under optimized conditions, the reaction time is selected to be 20min, and the absorbance of the reaction system at 652nm is measured. As shown in FIG. 18, N-CDs catalyze H at various concentrations2O2Oxidation of TMB produces a color change in the solution. With H2O2The concentration is increased, the absorbance of the system is also increased, and the concentration is increased in H2O2The absorbance reached a maximum at a concentration of 75. mu. mol/L, after which the absorbance of the solution was accompanied by H2O2The concentration increases and begins to decrease.
As shown in FIGS. 19-21, H2O2The concentration is in the range of 0.5-2.5 mu mol/L and 100-200 mu mol/L, and the absorbance and the concentration have good linear relation. H2O2When the concentration is 0.5-2.5 mu mol/L, the linear fitting equation is that y is 0.0684x +0.0178, R2＝0.9950。H2O2When the concentration is 100-200 mu mol/L, the linear fitting equation is that y is-5.28 x 10-4+0.5012，R20.9934. The detection limit (3. sigma./K) was 0.38. mu. mol/L.
5. Enzyme-linked immunosorbent assay for measuring hydrogen peroxide concentration
Reaction for 20min with microplate reader for determination of hydrogen peroxide concentration, as shown in FIGS. 22-24, H2O2The relationship between concentration and absorbance is similar to the curve measured by an ultraviolet spectrophotometer, but the OD value measured by an enzyme label instrument is larger. H2O2The concentration is in the range of 0.5-2.5 mu mol/L and 100-200 mu mol/L, and the absorbance and the concentration have good linear relation. H2O2When the concentration is 0.5-2.5 mu mol/L, the linear fitting equation is that y is 0.0676x +0.101, R2＝0.9935。H2O2When the concentration is 100-200 mu mol/L, the linear fitting equation is that y is-0.0006 x +0.5904, R20.9921. The detection limit (3. sigma./K) was 0.077. mu. mol/L.
TABLE 1 detection of different nanomaterials H2O2Comparison of
6. Reaction for 15min enzyme-linked immunosorbent assay to determine the concentration of hydrogen peroxide
To test whether the reaction time would be on H2O2And OD values, and a linear relationship of 15min was tested for reaction time. As can be seen from FIGS. 25 and 26, at this reaction time, the concentration of H was low2O2The linear range of (a) increases. H2O2When the concentration is 0.25-5 mu mol/L, the linear fitting equation is that y is 0.02930x +0.0646, R2＝0.9911。
7. Reaction 25min enzyme-linked immunosorbent assay for measuring hydrogen peroxide concentration
To test whether the reaction time would be on H2O2And OD values, and thus a linear relationship with a reaction time of 25min was tested. As can be seen from FIGS. 27 and 28, at this reaction time, the H concentration was higher2O2The linear range of (a) is increased but the linearity of the entire curve is not high. H2O2When the concentration is 50-200 mu mol/L, the linear fitting equation is that y is-0.0007 x +0.679, R2＝0.9746。
In summary, the N-CDs prepared by using citric acid and urea as raw materials and adopting a simple microwave method have the characteristic of hydrogen peroxide mimic enzyme, and can be used for catalyzing H by using tetramethyl benzidine (TMB) as a color developing agent2O2Performing color reaction with the above extract, performing spectrophotometry at N-CDs 150 μ g/ml, TMB 500 μmol/L, pH 3.2.2 at 35 deg.C for 20min, and performing H reaction2O2Has good linear relation with the absorbance within the range of 0.5-2.5 mu mol/L and 100-200 mu mol/L. Using a microplate reader for quantification, the detection method is also for H2O2Has good linear relation with OD value in the range of 0.5-2.5 mu mol/L and 100-200 mu mol/L, and the detection limit is 0.077 mu mol/L. The method is simple to operate and low in cost, can realize rapid detection of large samples, and also meets the detection requirement of micro-concentration. Quantifying by adopting an enzyme-labeling instrument under the conditions of 150 mu g/ml N-CDs, 500 mu mol/L TMB, pH3.2, reaction temperature of 35 ℃ and reaction time of 15min to obtain H2O2The concentration is 0.25-5 mu mol/L, and the OD value has good linear relation. H of the detection method when the reaction time is 25min2O2The fitted linearity of concentration and OD value is reduced, and the linear relation is not good.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.
Claims (3)
1. A method for detecting hydrogen peroxide is characterized by comprising the following steps:
preparation of a standard curve of S1: dividing the mixed solution of nitrogen-doped carbon quantum dots, ethanol solution of tetramethyl benzidine and acetate buffer solution with pH value of 3.2 into more than 3 parts, respectively adding hydrogen peroxide solutions with different concentrations, reacting at 35 ℃ for 15-25 min, transferring into a pore plate, placing into an enzyme labeling instrument for detection, and making a standard curve;
and S2, adding the hydrogen peroxide solution to be detected into the mixed solution with the same concentration as that in the step S1, reacting at 35 ℃ for 15-25 min, transferring into a pore plate, placing into an enzyme labeling instrument, detecting, and determining the concentration of the hydrogen peroxide solution to be detected.
2. The method of claim 1, wherein the nitrogen-doped carbon quantum dot is prepared by the following steps: the concentration of the citric acid in the mixed solution is 0.04-0.06 g/ml, and the concentration of the urea in the mixed solution is 0.02-0.04 g/ml.
3. The method of claim 1, wherein the method of fabricating the nitrogen-doped carbon quantum dot comprises the steps of: and (2) carrying out microwave treatment on the mixed solution of citric acid and urea in microwave for 2.5-4 min, adding ultrapure water for dissolving, centrifuging, taking supernatant, filtering with a 0.22-micron filter membrane, and drying the dialyzed solution in a 1000Kd dialysis bag to obtain the nitrogen-doped carbon quantum dot.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109781976A (en) * | 2018-12-27 | 2019-05-21 | 中国农业大学 | Fluorescence immune analysis method based on carbon quantum dot |
| CN110093158A (en) * | 2019-05-31 | 2019-08-06 | 西北大学 | A kind of fluorescent carbon quantum dot of N doping and its preparation method and application |
| CN110231297A (en) * | 2019-06-14 | 2019-09-13 | 西北师范大学 | The method for visualizing measurement content of hydrogen peroxide using doping type carbon quantum dot |
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| CN110093158A (en) * | 2019-05-31 | 2019-08-06 | 西北大学 | A kind of fluorescent carbon quantum dot of N doping and its preparation method and application |
| CN110231297A (en) * | 2019-06-14 | 2019-09-13 | 西北师范大学 | The method for visualizing measurement content of hydrogen peroxide using doping type carbon quantum dot |
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| CN114950530A (en) * | 2022-06-22 | 2022-08-30 | 广西大学 | Preparation method of nitrogen-doped eggshell nano enzyme with peroxidase-like activity |
| CN114950530B (en) * | 2022-06-22 | 2023-12-29 | 广西大学 | Preparation method of nitrogen-doped eggshell nanoenzyme with peroxidase-like activity |
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