CN115950866A - G-C3N 4/RB-based ratio fluorescence sensor and application thereof in carbendazim detection - Google Patents
G-C3N 4/RB-based ratio fluorescence sensor and application thereof in carbendazim detection Download PDFInfo
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- CN115950866A CN115950866A CN202211634791.4A CN202211634791A CN115950866A CN 115950866 A CN115950866 A CN 115950866A CN 202211634791 A CN202211634791 A CN 202211634791A CN 115950866 A CN115950866 A CN 115950866A
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Abstract
The invention discloses a method based on g-C 3 N 4 a/RB ratiometric fluorescent sensor consisting of g-C and its use in carbendazim detection 3 N 4 And a rhodamine B solution, wherein blue g-C is contained in the composite system 3 N 4 Fluorescence intensity ratio to rhodamine B I 450 /I 570 Is 3.5. Carbendazim molecules can act on g-C through electrostatic action 3 N 4 Surface enrichment, formation of pi-pi stacking, and causing electron transfer leading to g-C 3 N 4 The blue fluorescence of (a) is quenched, and Rhodamine B (RB) which emits orange-yellow fluorescence is used as an internal standard, the fluorescence color of the internal standard is not changed, so that blue fluorescence g-C is used 3 N 4 The ratio fluorescence sensor is prepared by taking orange-yellow fluorescent rhodamine B as a reference unit as a response unit, and an integrated intelligent mobile phone sensing platform is constructed by relying on a mature 3D printing technology, so that low-cost visual quantitative analysis of carbendazim pesticide residues is realized.
Description
Technical Field
The invention relates to a method based on g-C 3 N 4 A/RB ratio fluorescence sensor and application thereof in carbendazim detection belong to the fields of material synthesis, visual detection, food quality monitoring and the like.
Background
Carbendazim (carbendazim) is a common benzimidazole fungicide that has been widely used in agriculture, both before and after harvest, mainly on fruit, vegetable and cereal crops to control several fungal plant diseases such as spotting, mildew, mold, rot and focal spots. However, the carbendazim is low in degradation speed and large in residual quantity, has potential teratogenicity, reproductive toxicity and carcinogenicity, and is easy to accumulate in plants, so that the carbendazim enters a food chain and threatens human health. Therefore, the detection of the carbendazim, particularly the detection of the carbendazim in fruits, vegetables and drinking water, has very important practical significance.
At present, in the existing carbendazim detection technology, the preferred method is to combine liquid chromatography with different detector units for detection, and the national standard adopts reversed-phase high performance liquid chromatography to carry out detection and analysis on the carbendazim. The liquid chromatography usually needs complex and time-consuming sample pretreatment, and the application of the method in rapid trace detection is limited, so that an analysis method with stronger sensitivity, better selectivity and higher efficiency is urgently needed for rapid trace detection of carbendazim.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method based on g-C 3 N 4 the/RB ratio fluorescence sensor is used for high-sensitivity visual quantitative detection of carbendazim residue, and the sensing system can realize reliable, convenient and on-site carbendazim residue detection.
The fast visual ratio fluorescence sensor used as the core of the sensing system is a g-C sensor designed by integration 3 N 4 And rhodamine B to realize selective quantitative detection of carbendazim. Carbendazim is enriched to g-C by electrostatic action 3 N 4 On the nano-chip, pi-pi accumulation is formed, on the other hand, the oxidation-reduction potential of carbendazim is +1.052V and is positioned at g-C 3 N 4 between-1.09V (CB) and +1.56V (VB) of the valence band energy. Appropriate energy levelPromoting excited state electrons from g-C 3 N 4 Transfer of the nanoplatelets to the target analyte carbendazim molecule, resulting in g-C 3 N 4 The blue emission fluorescence quenches. While rhodamine B as a reference signal did not have any change in orange fluorescence at 570nm, thus resulting in a significant ratiometric fluorescent color change from blue to pink to orange. The detection limit of the carbendazim can be as low as 5.89nM. In addition, the ratio fluorescence sensing system has been successfully applied to carbendazim detection in practical samples, and a new method is provided for constructing a visual quantitative system for detecting trace hazards.
The invention is based on g-C 3 N 4 a/RB ratio fluorescence sensor consisting of g-C 3 N 4 And RB solution, wherein g-C in the composite system 3 N 4 The fluorescence intensity ratio to RB was 3.5.
The invention is based on g-C 3 N 4 The preparation method of the/RB ratio fluorescence sensor comprises the following steps:
step 1: g-C 3 N 4 Preparation of nanosheets
Putting 8-10 g of melamine powder into an alumina crucible with a cover, heating to 400-450 ℃ at a heating rate of 3 ℃ rise per minute, keeping the temperature at 400-450 ℃ for 1.5-2 hours, naturally cooling to room temperature to obtain a yellow blocky compound, grinding the yellow blocky compound into powder by using a ball mill, dispersing the yellow powder (0.8-0.1 g) into deionized water (80-100 ml) for ultrasonic treatment, centrifuging the obtained suspension to obtain a supernatant fluid, namely g-C 3 N 4 A nanosheet solution.
And 2, step: preparation of ratiometric fluorescent nano-sensing system
80-100 mu L of 1.0mg/mLg-C obtained in the step 1 3 N 4 Adding the nanosheet solution into 15-20 mu L of 1mM rhodamine B solution, diluting the nanosheet solution to 1.5-2 mL by using pure water in a 3mL cuvette, and fully mixing to obtain the ratiometric fluorescent nano sensing system.
The invention is based on g-C 3 N 4 Use of a/RB ratio fluorescence sensor for producing a detection reagent for visualizingAnd quantitatively detecting carbendazim residue.
The core of the visual quantitative detection of carbendazim in the sample is the designed g-C 3 N 4 (ii) ratio of/RB fluorescence sensing system. Carbendazim and g-C 3 N 4 (ii) a ratio of RB to g-C enrichment of carbendazim by electrostatic interaction 3 N 4 On the nano-chip, pi-pi accumulation is formed, and proper energy level promotes excited state electrons from g-C 3 N 4 Transfer of the nanoplatelets to the target analyte carbendazim molecule, resulting in g-C 3 N 4 The blue emission fluorescence is quenched, the RB as an internal standard has no change of orange-red fluorescence, and the color of the system continuously changes from blue to orange-red, so that the visual detection of the ratiometric fluorescence sensing system on the carbendazim is realized, and the content of the carbendazim in the sample is detected through the fluorescence change degree in the ratiometric fluorescence sensing system.
The detection method specifically comprises the following steps:
detection of carbendazim by ratiometric fluorescent probe solution
Dripping the ratio fluorescence nanoprobe solution into a quartz colorimetric tube of 3mL, sequentially adding 2-3 mu L of carbendazim solution with different concentrations, and mixing and shaking for 15-20 minutes. Obvious fluorescent color change can be observed under an ultraviolet lamp, and the visual detection of the carbendazim is realized. And recording the fluorescence spectrum of the solution in the range of 400-700 nm by using 320nm exciting light, and realizing the quantitative detection of the carbendazim by establishing the relation between the change of the fluorescence peak intensity ratio and the concentration of the carbendazim.
The principle of using the fluorescence sensor to detect carbendazim in the invention is based on a fluorescence quenching strategy, particularly, the prepared g-C 3 N 4 The positive electricity of the nano-sheets and the electrostatic action of the carbendazim with negative electricity cause enrichment, which results in pi bonds and g-C of the carbendazim 3 N 4 The pi bonds of the nano sheets generate pi-pi stacking, and on the other hand, the oxidation-reduction potential of the carbendazim is +1.052V and is positioned at g-C 3 N 4 between-1.09V (CB) and +1.56V (VB) of the valence band energy. Appropriate energy levels promote excited state electrons from g-C 3 N 4 The nanoplates are transferred to target analyte carbendazim molecules, therebyResult in g-C 3 N 4 The blue emission fluorescence was quenched and the reaction was complete within 20 minutes. Based on blue fluorescence quenching and orange-red fluorescence invariance, the sensing system shows the conversion from blue fluorescence to orange-red fluorescence under an ultraviolet lamp, and the visual detection of the carbendazim is realized; the quantitative detection of the carbendazim is realized by establishing the relation between the ratio fluorescence intensity and the concentration of the carbendazim.
Compared with the prior detection technology, the invention has the beneficial effects that:
1. the ratio fluorescence sensing system disclosed by the invention is used for detecting carbendazim, shows a better visual detection effect compared with other monochromatic fluorescence detection, effectively avoids the problem of instability of monochromatic fluorescence intensity, and realizes visual detection.
2. The detection limit of the fluorescence spectrometer for the carbendazim is 5.89nM and is lower than the residual allowable limit of the carbendazim.
3. Blue g-C prepared by the invention 3 N 4 And a fluorescence quenching system of the RB ratio sensor has good selectivity and sensitivity to carbendazim, can effectively avoid the interference of other impurities, and has quick response.
Drawings
FIG. 1 is g-C 3 N 4 Transmission electron microscopy of nanoplatelets.
FIG. 2 is g-C 3 N 4 XRD pattern of nanoplatelets.
FIG. 3 is g-C 3 N 4 Infrared spectrum of the nanosheet.
FIG. 4 is g-C 3 N 4 And (3) a system x-ray photoelectron energy spectrum of the nanosheet.
FIG. 5 Zeta potential diagrams of different samples.
FIG. 6 is a graph of optimal excitation wavelengths for a ratiometric fluorescence sensing system.
FIG. 7 is a graph of optimal response time for a ratiometric fluorescent sensing system. (A) g-C 3 N 4 Kinetic response to carbendazim; the reaction (B) was stable within 20 minutes.
FIG. 8 is a graph of pH optimum for a ratiometric fluorescence sensing system. (A) When the pH value is 4 to 10, the change of the fluorescence spectrum of the carbendazim proportional sensing system is added; (B) The pH value is plotted against the influence of a proportional fluorescent probe on the detection of carbendazim.
FIG. 9 is a ratio of optimal fluorescence intensities for the ratiometric fluorescence sensing system. After adding different concentrations of carbendazim, the fluorescence intensity ratios (I) were varied 450 /I 570 ) The fluorescence intensity of the constructed ratio sensing system changes. Fluorescence intensity ratio (I) 450 /I 570 ) FIG. 3 is a drawing (A); FIG. 3.5 is the drawing; FIG. 4 is a drawing (C).
FIG. 10 (A) is a graph showing the different concentrations of carbendazim versus g-C 3 N 4 Fluorescence spectrum and color change chart of/RB mixed system. The fluorescence color of the solution gradually changes from blue to orange-red along with the increase of the concentration of the carbendazim; FIG. 10 (B) is a graph showing the relationship between the fluorescence intensity and the concentration of carbendazim.
FIG. 11 (A) shows carbendazim g-C at different concentrations 3 N 4 a/RB mixed system smartphone color identification schematic diagram; FIG. 11B is a graph showing the relationship between the color change of the fluorescent probe solution (red channel/blue channel) and the concentration of carbendazim, and there is a linear relationship between the color change of the fluorescent probe solution (red channel/blue channel) and the concentration of carbendazim.
FIG. 12 (A, B, C) is a graph of fluorescence probe selectivity and interference.
Detailed Description
The technical scheme of the invention is further explained by combining the drawings and the specific embodiments as follows:
example 1: preparation of ratiometric fluorescent nanosensors
1、g-C 3 N 4 Preparation of nanosheets
Putting 8-10 g of melamine powder into an alumina crucible with a cover, heating to 400-450 ℃ at a heating rate of 3 ℃ rise per minute, keeping the temperature at 400-450 ℃ for 1.5-2 hours, and naturally cooling to room temperature. Grinding the obtained yellow block-shaped compound into powder by a ball mill, dispersing the yellow powder (0.8-0.1 g) into deionized water (80-100 ml) for ultrasonic treatment, centrifuging the obtained suspension, and obtaining supernatant fluid which is g-C 3 N 4 A nanosheet solution.
Step 2: preparation of ratiometric fluorescent nano-sensing system
Will 80E-mail100 μ L of 1.0mg/mLg-C obtained in step 1 3 N 4 Adding the solution into 15-20 mu L of 1mM rhodamine B solution, diluting the solution to 1.5-2 mL by pure water in a 3mL cuvette, and fully mixing to obtain a ratio fluorescence nano sensing system.
Example 2: ratiometric fluorescence sensing mechanisms, component characterization, and optimization of detection conditions
1. The ratiometric fluorescence sensing system has obvious dual emission peaks under 365nm ultraviolet excitation. After introduction of carbendazim, g-C 3 N 4 Blue fluorescence at 450nm can be quenched by Photoinduced Electron Transfer (PET) within 20 minutes. And the rhodamine B is used as a reference signal, and the orange-red fluorescence of the rhodamine B at 570nm does not change, so that the obvious color change from blue to pink to orange can be caused, the rapid visual detection of the ratiometric fluorescence sensing system on the carbendazim can be realized, and the content of the carbendazim in the sample can be detected through the fluorescence change degree in the ratiometric fluorescence system.
2. Ratiometric fluorescence sensing system component characterization
Considering that the detection sensitivity of the probe to the analyte is related to its own properties, the structural features and spectral characteristics of ratiometric fluorescent nanoprobes were studied using FT-IR, UV-vis and fluorescence spectroscopy, respectively. In addition, TEM, EDX and XPS were used to determine the morphology of ratiometric fluorescent nanoprobes and their elemental composition.
3. Excitation wavelength, reaction time, pH and g-C 3 N 4 Effect of/RB fluorescence intensity ratio on comparative fluorescent probes
Excitation wavelength, pH and g-C 3 N 4 the/RB fluorescence intensity ratio has certain influence on the fluorescence intensity and the detection effect of the ratiometric fluorescent probe. As can be seen from FIG. 6, g-C at different excitation wavelengths 3 N 4 The fluorescence intensity of blue is different from that of rhodamine B orange red, which shows that the fluorescence intensity of the fluorescence probe with the excitation wavelength and the contrast ratio has influence. In general, 320nm was selected as the optimal excitation wavelength for the probe of the present invention.
The reaction time is shown in FIG. 7, and the fluorescence is not changed when the reaction time is longer than 20 minutes. g-C under alkaline conditions 3 N 4 The blue fluorescence intensity is weakened, no fluorescence change is caused after the carbendazim is added, the fluorescence intensity is increased when the pH value is gradually reduced, and the fluorescence intensity change is most obvious when the pH value is equal to 6 after the carbendazim is added. Considering we chose 20 minutes as the optimal reaction time and 6 as the optimal pH for the assay.
g-C 3 N 4 And the fluorescence intensity of rhodamine B plays an important role in comparison with the detection effect of carbendazim. As shown in FIG. 9, the emission intensity ratio (I) at 365nm UV light 450 /I 570 ) At the 3.5 adjustment, a clear fluorescent color change from blue to pink was seen in the sensor solution after carbendazim addition.
Example 3:
1. detection of carbendazim by ratiometric fluorescent probe solution
The ratiometric fluorescent nanoprobe solution prepared in example 1 is dropped into a quartz colorimetric tube of 3mL, 2 to 3 μ L of carbendazim solutions with different concentrations are sequentially added, and the mixture is mixed and shaken uniformly. Obvious fluorescent color change can be observed under an ultraviolet lamp, and the visual detection of the carbendazim is realized. And recording the fluorescence spectrum of the solution in the range of 400-700 nm by using 320nm exciting light, and realizing the quantitative detection of the carbendazim by establishing the relation between the change of the fluorescence peak intensity ratio and the concentration of the carbendazim.
2. Drawing of standard curve
To g-C 3 N 4 The fluorescence intensity is tested after mixing carbendazim solutions with different concentrations are respectively added into a mixed system of the RB ratio probe, the result shows that the blue fluorescence emission peak at 450nm is gradually weakened, the fluorescence emission peak at 570nm is almost unchanged, and the fluorescence intensity ratio (I) is established 450 /I 570 ) And the relation between the concentration of the carbendazim can realize quantitative detection of the carbendazim. When the exciting light is 320nm, the fluorescence spectrum of the mixed system in the wavelength range of 400-700 nm is recorded. FIG. 10 shows the relationship between fluorescence intensity and carbendazim concentration, with a linear relationship between the change in the ratio of fluorescence intensity, wherein the abscissa is the concentration of carbendazim and the ordinate is the ratio of fluorescence intensity at 450nm and 570 nm.
Example 4:
1、g-C 3 N 4 preparation of nanosheets
The procedure for this step was the same as in example 1.
2. Preparation of ratiometric fluorescent nanosensors
The procedure for this step is the same as in example 1.
3. Construction of detection platform of smart phone
The smart phone detection platform comprises a colorimetric tank, a darkroom, a small UV lamp support, a filter (400-1100 nm) and a smart phone (OPPOA 53).
5. Probe solution detection of carbendazim
Adding the fluorescent nano probe solution into a cuvette, then adding the carbendazim solution with different concentrations, and shaking and mixing for 15-20 minutes. And acquiring a fluorescence image through software of the smart phone in a dark environment, further decomposing the fluorescence image into RGB values, and completing quantitative detection by installing a color recognition Application (APP).
6. Drawing of standard curve
A ratio fluorescence sensing cuvette dropwise added with carbendazim solutions of different concentrations is placed in a detection platform, and a fluorescence image is obtained through a smart phone by taking a 365nm ultraviolet lamp as a light source in a dark environment. Further decomposed into RGB values, and the inset 11 shows that the RGB values are linear with the carbendazim concentration, where the abscissa is the carbendazim concentration and the ordinate is the RGB ratio.
Example 5:
1、g-C 3 N 4 preparation of nanosheets
The procedure for this step was the same as in example 1.
2. Preparation of ratiometric fluorescent nanosensors
The procedure for this step is the same as in example 1.
4. Fluorescence nanoprobe selectivity and interference testing
Adding 140-150 nM Trichlorfon (Trichlorofon), dimethoate (Dimethoate), acephate (Acephate), glyphosate ethyl paraoxon (Glyphosate), chlorothalonil (Chlorothalonil) and inorganic ions (Zn) into a fluorescent nano probe system 2+ ,K + ,Ca 2+, Na + ,Mg 2+ ,Al 3+ ) The result shows that the fluorescence intensity ratio has no obvious change, and the blue fluorescence is quenched and the fluorescence intensity ratio is obviously changed by continuously adding 140-150 nM carbendazim, as shown in figure 12. The results show that the system has good selectivity and interference resistance to carbendazim.
Claims (9)
1. Based on g-C 3 N 4 a/RB ratio fluorescence sensor, characterized in that:
the ratiometric fluorescence sensor consists of g-C 3 N 4 And a rhodamine B solution, wherein blue g-C is contained in the composite system 3 N 4 Fluorescence intensity ratio to rhodamine B I 450 /I 570 Is 3.5.
2. A composition according to claim 1 based on g-C 3 N 4 The preparation method of the/RB ratio fluorescence sensor is characterized by comprising the following steps:
step 1: g-C 3 N 4 Preparation of nanosheets
Putting 8-10 g of melamine powder into an alumina crucible with a cover, heating to 400-450 ℃ at a heating rate of 3 ℃ rising per minute, keeping at 400-450 ℃ for 1.5-2 hours, naturally cooling to room temperature, grinding the obtained yellow blocky compound into powder by using a ball mill, dispersing the yellow powder into deionized water, performing ultrasonic treatment, centrifuging the obtained suspension, and obtaining the supernatant which is g-C 3 N 4 A nanosheet solution;
and 2, step: preparation of ratiometric fluorescent nano sensing system
80-100 mu L of 1.0mg/mLg-C obtained in the step 1 3 N 4 Adding the nanosheet solution into 15-20 mu L of 1mM rhodamine B solution, diluting the nanosheet solution to 1.5-2 mL by using pure water in a 3mL cuvette, and fully mixing to obtain the ratiometric fluorescent nano sensing system.
3. The method of claim 2, wherein:
in the step 1, the grinding time of the ball mill is more than or equal to 6 hours.
4. The production method according to claim 2, characterized in that:
in the step 1, 0.8-0.1 g of yellow powder is dispersed into 80-100 mL of deionized water, the ultrasonic time is 8-10 hours, and the obtained suspension is centrifuged for 15-20 minutes.
5. A composition according to claim 1 based on g-C 3 N 4 The application of the/RB ratio fluorescence sensor in the preparation of detection reagents is characterized in that: the detection reagent is used for visually and quantitatively detecting carbendazim.
6. Use according to claim 5, characterized in that:
the ratio fluorescence sensor is used as a detection reagent and is dripped into a 3mL quartz colorimetric tube, 2-3 mu L of a sample to be detected is added, the mixture is mixed and vibrated for 15-20 minutes, obvious fluorescence color change can be observed under an ultraviolet lamp, and the visual detection of the carbendazim is realized.
7. Use according to claim 5, characterized in that:
the fluorescence sensor with the ratio is used as a detection reagent and is dripped into a quartz colorimetric tube with 3mL, 2-3 mu L of carbendazim solutions with different concentrations are sequentially added, mixed and vibrated for 15-20 minutes, the fluorescence spectrum of the solution in the range of 400-700 nm is recorded by 320nm luminescence, and the quantitative detection of the carbendazim is realized by establishing the relation between the change of the fluorescence peak intensity ratio and the concentration of the carbendazim.
8. Use according to claim 5, characterized in that:
under an ultraviolet light source, images and RGB values are obtained by software, and quantitative detection is realized by establishing the relation between the ratio of R to B and the concentration of carbendazim.
9. Use according to claim 5, characterized in that:
the detection limit of the detection reagent on carbendazim is 5.89nM.
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