A probe for rapid, high-sensitivity colorimetric and fluorescent dual-mode detection of hydrogen peroxide, and a preparation method and use thereof

Abstract

The application provides a probe for rapid and high-sensitivity colorimetric and fluorescent dual-mode detection of hydrogen peroxide, a preparation method and use, and benefits from the unique design of the carboxyl on the probe, which can consume the intermediate product hydroxyl anion produced in the reaction of the probe with hydrogen peroxide, so that the reaction proceeds in the forward direction, accelerates the generation of product molecules, and the carboxyl makes the probe have a self-accelerating effect, and finally realizes rapid and high-sensitivity dual-mode detection of hydrogen peroxide. The solution color of the probe changes from light yellow to golden yellow within 5s of contact with hydrogen peroxide, and the detection limit is 236.84nM. At the same time, under the excitation of 468nm blue light, the fluorescence rapidly changes from green to yellow-green, and the detection limit is as low as 3.36nM. The probe has an ultrafast reaction speed and an excellent detection limit, and the detection process does not require complex analysis equipment, realizing colorimetric and fluorescent dual-mode trace detection of hydrogen peroxide. Therefore, the design of the probe can provide effective technical support for the field of trace detection of non-standard explosives in actual environment.

Description

Technical Field

[0001] This invention belongs to the field of analytical detection and provides a rapid, highly sensitive probe and detection method for dual-mode detection of hydrogen peroxide using colorimetry and fluorescence. This probe has a fast response time and a low detection limit, enabling rapid and efficient dual-mode detection of hydrogen peroxide. Background Technology

[0002] Hydrogen peroxide, due to its strong oxidizing properties, is the preferred raw material for preparing peroxide explosives. At the same time, it is also a decomposition product of peroxide explosives. Therefore, rapid and ultrasensitive on-site identification and detection of hydrogen peroxide is of great practical significance.

[0003] Currently, a series of analytical techniques for the detection of hydrogen peroxide have been developed, including mass spectrometry (Analytical Methods 2010, 2, 897-900), high-performance liquid chromatography (High-performance liquid chromatography) (Food Chemistry 2019, 283, 431-436), surface-enhanced Raman spectroscopy (AIChE Journal 2017, 63, 3994-4002), electrochemical detection (Process Biochemistry 2022, 115, 57-69), and fluorescence detection. Among these, fluorescence detection has attracted widespread attention due to its high sensitivity, fast reaction speed, strong specificity, visualized detection signal, and ease of operation. To date, researchers have designed and synthesized a large number of fluorescent probes for the detection of hydrogen peroxide. For example, Professor Song Xiangzhi's research group at Central South University designed and synthesized a 1,3-bis(bispyridine-2-imino)isoindoline-4-ol fluorescent probe to achieve fluorescence detection of hydrogen peroxide (Sensors and Actuators B: Chemical 2018, 255, 1160-1165); Professor Shang Luqing's research group at Nankai University designed and synthesized a dual-mechanism fluorescent probe using intramolecular charge transfer-fluorescence resonance energy transfer with coumarin as the energy donor and naphthalimide as the energy acceptor to achieve ratiometric fluorescence detection of hydrogen peroxide (Chemistry-An Asian Journal). (2017, 12, 3187-3194); The Zang Ling research group at the University of Utah designed and synthesized a fluorescent activating probe molecule of N-hexane-4-borate pinacol ester-1,8-naphthalimide. Using naphthalimide as the fluorophore and borate ester as the recognition group, they achieved fluorescence detection of hydrogen peroxide vapor. This probe rapidly produces yellow-green fluorescence upon contact with hydrogen peroxide vapor (Chemical Communications 2013, 49, 11779-11781); Chinese patent CN 116143814 discloses a curcumin-based fluorescent probe for detecting hydrogen peroxide. After the addition of hydrogen peroxide, the fluorescence color of the solution changes from orange to colorless under 365nm ultraviolet light. These fluorescence detection technologies, as single-channel signal sensing technologies, are easily affected by fluorescent background and complex fluorescent substances commonly found in daily life, and their detection accuracy needs improvement.

[0004] A common and efficient strategy to address this problem is to construct a dual-mode visualization detection platform, generating two independent detection signals to effectively reduce the influence and potential interference from the external environment. Currently, researchers have developed many organic small molecule fluorescent probes with integrated colorimetric and fluorescence dual-mode detection. For example, Professor Zhu Baocun's group at Jinan University designed and synthesized a fluorescent probe with naphthalimide as the fluorophore and pinacol boronic acid as the recognition group, capable of colorimetric fluorescence dual-mode detection of hydrogen peroxide. After adding hydrogen peroxide for 10 minutes, the fluorescence changed from blue-violet to green, with a detection limit of 4 μM. Simultaneously, the solution color changed from colorless to yellow, with a detection limit of 50 μM (RSC Advances 2014, 4, 16055-16061). The research group of Li Haitao at Hunan Normal University designed and synthesized a colorimetric-near-infrared fluorescence dual-mode probe for detecting hydrogen peroxide, using dicyanomethylene-4H-pyran as the fluorophore and pinacol borate ester as the recognition unit. After 30 min of hydrogen peroxide addition, the fluorescence significantly increased, emitting red fluorescence, with a detection limit of 70 nM, and the solution color changed from yellow to purple (Sensors and Actuators BChemical 2019, 280, 120-128). The research group of Lin Weiying at Jinan University designed and synthesized a lysosome-targeted two-photon fluorescent probe, using 1,8-naphthalimide as the fluorophore and borate as the reaction site. After 60 s of hydrogen peroxide addition, the fluorescence increased, emitting green fluorescence, with a detection limit of 1.21 μM, and the solution color changed from colorless to yellow (Biosensors and Bioelectronics 2016, 79, 237-243). However, most reported colorimetric-fluorescence dual-mode systems for detecting hydrogen peroxide currently suffer from slow reaction rates and low sensitivity. Therefore, it is particularly important to develop a dual-mode detection probe for hydrogen peroxide that is fast-responding, has low sensitivity, and is portable, using both colorimetric and fluorescent methods.

[0005] This invention, starting from the structure of the probe molecule itself, uses a carboxyl group as a self-accelerating group, 1,8-naphthalimide as a fluorophore, and pinacol borate as a recognition group to develop a rapid and highly sensitive colorimetric and fluorescence dual-mode probe for the detection of hydrogen peroxide. The detection principle is that the pinacol borate on the probe reacts with hydrogen peroxide in the presence of tetrabutylammonium hydroxide to generate 4-hydroxyphenylboronic acid pinacol ester and hydroxide anion intermediates. The carboxyl group on the probe consumes the hydroxide anion intermediate, causing the reaction to proceed in the forward direction. The design of the carboxyl group on the probe acts as a self-accelerator, thus rapidly generating a phenol salt product that exhibits both yellow color and yellow-green fluorescence, ultimately achieving colorimetric and fluorescence dual-mode detection of hydrogen peroxide. This reagent is characterized by its rapid speed, high sensitivity, simple operation, and ease of widespread application. Summary of the Invention

[0006] The purpose of this invention is to provide a rapid, highly sensitive colorimetric and fluorescence dual-mode probe for the detection of hydrogen peroxide, along with its preparation method and applications. The probe's chemical name is N-hexanoic acid-4-borate pinacol ester-1,8-naphthalimide. The design of the carboxyl group on the probe allows it to consume the intermediate hydroxide anion generated during the reaction between the probe and hydrogen peroxide, thereby accelerating product formation. Within 5 seconds of contact with hydrogen peroxide, the solution color rapidly changes from pale yellow to gold, with a detection limit of 236.84 nM. Simultaneously, under 468 nm blue light excitation, the fluorescence rapidly changes from green to yellow-green, with a detection limit as low as 3.36 nM. This probe overcomes the shortcomings of slow molecular response and low sensitivity in hydrogen peroxide detection probes, providing effective technical support for the field of trace detection of non-standard explosives.

[0007] The present invention discloses a rapid and highly sensitive colorimetric and fluorescence dual-mode detection probe for hydrogen peroxide, the chemical structural formula of which is (1):

[0008]

[0009] The chemical name of this probe is: N-hexanoic acid-4-boronic acid pinacol ester-1,8-naphthalimide.

[0010] The method for preparing the probe for rapid, highly sensitive colorimetric and fluorescence dual-mode detection of hydrogen peroxide comprises the following steps:

[0011] a) Dissolve 4-bromo-1,8-naphthalenedicarboxylic anhydride in ethanol, then add 6-aminohexanoic acid, set the temperature to 90℃, react for 12 h, cool to room temperature, precipitate and wash with ice water, dry, and purify by column chromatography to obtain the intermediate product N-hexanoic acid-4-bromo-1,8-naphthalenedicarboxylic anhydride, wherein the molar ratio of 4-bromo-1,8-naphthalenedicarboxylic anhydride to 6-aminohexanoic acid is 4:5;

[0012] b. The intermediate product N-hexanoic acid-4-bromo-1,8-naphthylimide, pinacol diborate, and potassium acetate obtained in step a were dissolved in the solvent 1,4-dioxane. After purging with nitrogen for 30 min, 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride was added. The reaction was carried out overnight at 90 °C under a nitrogen atmosphere. After cooling to room temperature, the mixture was diluted with dichloromethane, washed three times with deionized water and brine, dried over anhydrous magnesium sulfate, filtered, and purified by column chromatography to obtain the probe N-hexanoic acid-4-borate pinacol dichloride-1,8-naphthylimide. The molar ratio of the intermediate product N-hexanoic acid-4-bromo-1,8-naphthylimide to pinacol diborate, potassium acetate, and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride was 2:3:6:3 / 25.

[0013] The application of the aforementioned probe for rapid, highly sensitive colorimetric and fluorescence dual-mode detection of hydrogen peroxide is carried out according to the following steps:

[0014] a. Dissolve the probe N-hexanoic acid-4-borate pinacol ester-1,8-naphthalimide in ethanol solvent to prepare a probe solution with a concentration of 1 mM, and then add 1 mM tetrabutylammonium hydroxide to obtain the detection reagent.

[0015] b. After adding hydrogen peroxide to the test reagent obtained in step a, the solution color rapidly changes from light yellow to gold within 5 seconds. Under 468nm blue light excitation, the fluorescence changes from green to yellow-green.

[0016] When the hydrogen peroxide concentration is 0-350 μM, the absorption peak intensity at 454 nm is linearly related to the hydrogen peroxide concentration, and the detection limit is 236.84 nM; when the hydrogen peroxide concentration is 0-80 μM, the fluorescence emission peak intensity at 558 nm is linearly related to the hydrogen peroxide concentration, and the detection limit is 3.36 nM.

[0017] This invention discloses a rapid, highly sensitive colorimetric and fluorescence dual-mode detection probe for hydrogen peroxide, its preparation method, and its applications. The detection principle of this probe is as follows: the recognition site of the organic small molecule fluorescent probe N-hexanoic acid-4-boronate-1,8-naphthalimide is borate pinacol ester. Under the action of tetrabutylammonium hydroxide catalyst, it can be oxidized to phenol by hydrogen peroxide, resulting in intramolecular charge transfer of the fluorophore 1,8-naphthalimide. The unique design of the carboxyl group on the probe of this invention can consume the hydroxide anions generated during the reaction with hydrogen peroxide, accelerating the formation of product molecules and achieving a self-accelerating process. This effectively promotes the reaction process, thereby greatly improving the response speed and sensitivity of the probe for hydrogen peroxide detection. It achieves a rapid change in solution color from light yellow to gold, and simultaneously, under 468nm blue light excitation, the fluorescence changes from green to yellow-green. This solves the current problems of slow response speed and low sensitivity for hydrogen peroxide detection.

[0018] This invention discloses a rapid, highly sensitive colorimetric and fluorescence dual-mode detection probe for hydrogen peroxide, its preparation method, and its applications. The synthetic route for this probe, N-hexanoic acid-4-borate pinacol ester-1,8-naphthalimide, is as follows:

[0019]

[0020] The present invention discloses a method for preparing a rapid, highly sensitive colorimetric and fluorescence dual-mode probe for detecting hydrogen peroxide, characterized by the following steps:

[0021] a) Dissolve 4-bromo-1,8-naphthalenedicarboxylic anhydride in ethanol, then add 6-aminohexanoic acid, set the temperature to 90℃, react for 12 h, cool to room temperature, precipitate and wash with ice water, dry, and purify by column chromatography to obtain the intermediate product N-hexanoic acid-4-bromo-1,8-naphthalenedicarboxylic anhydride, wherein the molar ratio of 4-bromo-1,8-naphthalenedicarboxylic anhydride to 6-aminohexanoic acid is 4:5;

[0022] b. The intermediate product N-hexanoic acid-4-bromo-1,8-naphthylimide, pinacol diborate, and potassium acetate obtained in step a were dissolved in the solvent 1,4-dioxane. After purging with nitrogen for 30 min, 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride was added. The reaction was carried out overnight at 90 °C under a nitrogen atmosphere. After cooling to room temperature, the mixture was diluted with dichloromethane, washed three times with deionized water and brine, dried over anhydrous magnesium sulfate, filtered, and purified by column chromatography to obtain the probe N-hexanoic acid-4-borate pinacol dichloride-1,8-naphthylimide. The molar ratio of the intermediate product N-hexanoic acid-4-bromo-1,8-naphthylimide to pinacol diborate, potassium acetate, and 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride was 2:3:6:3 / 25.

[0023] The present invention discloses a rapid, highly sensitive colorimetric and fluorescence dual-mode detection probe and detection method for hydrogen peroxide. The specific method for hydrogen peroxide performance testing is as follows:

[0024] Measure hydrogen peroxide and dilute it in deionized water to prepare aqueous solutions of hydrogen peroxide with concentrations of 0 mM, 0.2 mM, 0.4 mM, 0.6 mM, 0.8 mM, 1.0 mM, 1.4 mM, 1.6 mM, 1.8 mM, 2.0 mM, 2.5 mM, 3.0 mM, 4.0 mM, 5.0 mM, 6.0 mM, 7.0 mM, and 8.0 mM.

[0025] 1.9 mL of probe solution was measured with a pipette and placed in a quartz cuvette. 100 μL of hydrogen peroxide aqueous solution of different concentrations was added. Colorimetric and fluorescence detection images before and after the reaction were recorded with a digital camera.

[0026] Alternatively, 1.9 mL of the probe solution can be pipetted into a quartz cuvette, and then 100 μL of hydrogen peroxide aqueous solutions of different concentrations can be added. A UV-Vis absorption spectrum scan can be performed using a UV-Vis spectrophotometer to measure the UV-Vis absorption spectra of the probe solution after detecting different concentrations of hydrogen peroxide. Then, a linear equation can be fitted with the maximum absorbance as the ordinate and the hydrogen peroxide concentration as the abscissa to determine the colorimetric detection limit of the probe solution.

[0027] Alternatively, 1.9 mL of probe solution can be pipetted into a quartz cuvette, and then 100 μL of hydrogen peroxide aqueous solution of different concentrations can be added. Fluorescence emission spectrum scanning can be performed using a fluorescence spectrometer to measure the fluorescence emission spectrum of the probe solution after detecting different concentrations of hydrogen peroxide. Then, a linear equation in one variable can be fitted with the fluorescence emission peak intensity as the ordinate and the hydrogen peroxide concentration as the abscissa to determine the fluorescence detection limit of the probe solution.

[0028] The present invention discloses a rapid and highly sensitive probe detection method for hydrogen peroxide using colorimetric and fluorescence dual-mode detection. This method is mainly used for the detection of hydrogen peroxide, a precursor of explosives, and solves the current problem of rapid, highly sensitive, and visual detection of hydrogen peroxide in field testing.

[0029] Compared with the prior art, the beneficial effects of the present invention are:

[0030] 1. The probe described in this invention has a unique carboxyl group design. The carboxyl group can consume the intermediate product hydroxide ions generated during the reaction between the probe and hydrogen peroxide, giving it the ability to accelerate itself. This enables fast detection of hydrogen peroxide, achieving dual-mode detection in the colorimetric and fluorescence channels within 5 seconds. At the same time, it has high detection sensitivity and a fluorescence detection limit as low as 3.36 nM.

[0031] 2. The probe described in this invention does not require complex analysis equipment and can be directly identified and detected with the naked eye;

[0032] 3. The probe described in this invention does not require any pretreatment of the analyte, is simple to operate, and is easy to promote and apply. It can fully achieve the purpose of low-cost colorimetric and fluorescence dual-mode detection of hydrogen peroxide. Attached Figure Description

[0033] Figure 1 This is the proton NMR spectrum of the probe of this invention;

[0034] Figure 2 This is the carbon NMR spectrum of the probe of this invention;

[0035] Figure 3 This is the mass spectrum of the probe of the present invention;

[0036] Figure 4 The UV-Vis absorption spectra of the probe in ethanol solvent with a concentration of 1 mM and tetrabutylammonium hydroxide with a concentration of 1 mM, before and after reaction with 250 μM hydrogen peroxide aqueous solution are shown. Figure 4 (Left) and fluorescence emission spectrum ( Figure 4 right);

[0037] Figure 5 The concentration of the probe in the ethanol solvent of this invention is 1 mM, and the concentration of tetrabutylammonium hydroxide is 1 mM. Figure 5The left image shows the UV-Vis absorption spectra obtained after reacting with hydrogen peroxide at concentrations of 0 μM to 400 μM. Figure 5 The right side shows a linear equation obtained by fitting hydrogen peroxide concentration as the x-axis and the absorbance at 454 nm as the y-axis.

[0038] Figure 6 The concentration of the probe in the ethanol solvent of this invention is 1 mM, and the concentration of tetrabutylammonium hydroxide is 1 mM. Figure 6 The left image shows the fluorescence emission spectra obtained after reaction with 0μM-250μM hydrogen peroxide, where... Figure 6 The right side shows a linear equation obtained by fitting hydrogen peroxide concentration as the x-axis and fluorescence emission peak intensity at 558 nm as the y-axis.

[0039] Figure 7 The colorimetric results obtained by digital camera after reacting the probe of this invention with 1 mM in ethanol solvent and 1 mM in tetrabutylammonium hydroxide with 0 μM-250 μM hydrogen peroxide are shown. Figure 7 (above) and fluorescence ( Figure 7 (Below) Image. Detailed Implementation

[0040] The present invention will be further described below through specific embodiments, but the invention is not limited to these embodiments.

[0041] Example 1

[0042] Probe preparation:

[0043] a) Dissolve 554.5 mg (2 mmol) of 4-bromo-1,8-naphthalenedicarboxylic anhydride in 30 mL of ethanol. While stirring, add 328.6 mg (2.5 mmol) of 6-aminohexanoic acid. Heat to 90 °C and react for 12 h. Cool to room temperature, pour the mixture into 100 mL of ice water, wash with deionized water, dry, and purify by column chromatography with dichloromethane:methanol at a volume ratio of 30:1 to obtain the intermediate product N-hexanoic acid-4-bromo-1,8-naphthalimide, which is a light yellow powder with a yield of 60%.

[0044] 1 H NMR (400MHz, CDCl3) δ (ppm): 8.66 (d, J = 7.3Hz, 1H), 8.57 (d, J = 8.5Hz, 1H), 8.41 (d, J = 7.9Hz, 1H), 8.04 (d, J = 7.9H z,1H),7.88-7.82(m,1H),4.20-4.14(m,1H),2.41-2.34(m,2H),1.74(dq,J=23.1,7.6Hz,4H),1.53-1.45(m,2H). 13C NMR(101MHz,CDCl3)δ(ppm):178.39,163.62,133.28,132.07,131.19,130.65,130 .27,129.02,128.09,123.10,122.23,40.26,33.65,27.66,26.49,24.35.HRMS:[M] + Calcd.for 390.23;Found 391.04;

[0045] b. The intermediate product obtained in step a, N-hexanoic acid-4-bromo-1,8-naphthalimide, 389.7 mg (1 mmol), pinacol diborate, 337.5 mg (1.5 mmol), and potassium acetate, 416.2 mg (3 mmol), were dissolved in 20 mL of 1,4-dioxane. After purging with nitrogen for 30 min, 26.7 mg (0.06 mmol) of 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride was added. The mixture was heated to 90 °C and reacted overnight under a nitrogen atmosphere. After cooling to room temperature, the mixture was diluted with 30 mL of dichloromethane, washed three times with deionized water and brine, dried over anhydrous magnesium sulfate, filtered, and purified by column chromatography with a dichloromethane:methanol solution at a volume ratio of 70:1 to obtain the probe N-hexanoic acid-4-borate pinacol diborate-1,8-naphthalimide, which was a brown solid with a yield of 78%.

[0046] 1 H NMR (400MHz, DMSO-d6) δ (ppm): 12.02 (s, 1H), 8.93 (d, J = 9.5Hz, 1H), 8.38 (dd, J = 10.5, 7.3Hz, 1H), 8.15 (d, J = 7.3Hz, 1H),7.86-7.80(m,1H),4.01-3.95(m,1H),2.22(t,J=7.2Hz,1H),1.66-1.51(m,1H),1.42(s,8H),1.37-1.31(m,4H). 13 C NMR(101MHz,DMSO-d6)δ(ppm):174.92,163.62,135.95,134.81,134.53,130.75,129.66,127.82,12 7.43(s),124.60,122.59,84.87,73.97,40.65,34.00,27.65,26.52,25.41,25.17,24.69.HRMS:[M] + Calcd.for 437.30;Found 438.21.

[0047] Example 2

[0048] Probe detection:

[0049] The probe prepared in step 1 was dissolved in ethanol solvent to prepare a probe solution with a concentration of 1 mM, and then 1 mM tetrabutylammonium hydroxide was added to obtain the detection reagent.

[0050] A 30% standard solution of hydrogen peroxide was diluted with water to obtain a 5 mM aqueous solution of hydrogen peroxide.

[0051] Measure 1.9 mL of the probe detection reagent into a quartz cuvette, then add 100 μL of a 5 mM hydrogen peroxide standard aqueous solution prepared by diluting hydrogen peroxide in deionized water. Within 5 seconds, the solution color rapidly changes from light yellow to gold. At the same time, under 468 nm blue light excitation, the solution fluorescence rapidly changes from green to yellow-green.

[0052] Example 4

[0053] The preparation of the probe and probe detection reagents was carried out according to the examples;

[0054] The probe prepared in step 1 was dissolved in ethanol solvent to prepare a probe solution with a concentration of 1 mM, and then 1 mM tetrabutylammonium hydroxide was added to obtain the detection reagent.

[0055] A standard solution of 30% hydrogen peroxide was diluted with water to obtain aqueous solutions of hydrogen peroxide with concentrations of 0 mM, 0.2 mM, 0.4 mM, 0.6 mM, 0.8 mM, 1.0 mM, 1.4 mM, 1.6 mM, 1.8 mM, 2.0 mM, 2.5 mM, 3.0 mM, 4.0 mM, 5.0 mM, 6.0 mM, 7.0 mM, and 8.0 mM.

[0056] Measure 1.9 mL of the probe detection reagent into a quartz cuvette, then add hydrogen peroxide to dilute it in deionized water to prepare 100 μL of hydrogen peroxide standard aqueous solutions of various concentrations. Perform UV-Vis absorption spectroscopy scanning using a UV-Vis spectrophotometer. By comparing the UV-Vis absorption spectra, it can be seen that the absorption peak intensity at 454 nm has a linear relationship with the hydrogen peroxide concentration from 0 μM to 350 μM, and the detection limit is 236.84 nM.

[0057] Example 5

[0058] The probe prepared in step 1 was dissolved in ethanol solvent to prepare a probe solution with a concentration of 1 mM, and then 1 mM tetrabutylammonium hydroxide was added to obtain the detection reagent.

[0059] A standard solution of 30% hydrogen peroxide was diluted with water to obtain aqueous solutions of hydrogen peroxide at concentrations of 0 mM, 0.2 mM, 0.4 mM, 0.6 mM, 0.8 mM, 1.0 mM, 1.4 mM, 1.6 mM, 1.8 mM, 2.0 mM, 2.5 mM, 3.0 mM, 4.0 mM, and 5.0 mM.

[0060] Measure 1.9 mL of the probe detection reagent into a quartz cuvette, then add hydrogen peroxide to dilute it in deionized water to prepare 100 μL of hydrogen peroxide standard aqueous solutions of various concentrations. Perform fluorescence emission spectroscopy scanning using a fluorescence spectrometer. By comparing the fluorescence spectra, it can be seen that the intensity of the fluorescence emission peak at 558 nm is linearly related to the hydrogen peroxide concentration from 0 μM to 80 μM, and the detection limit is 3.36 nM.

[0061] Example 6

[0062] The probe prepared in step 1 was dissolved in ethanol solvent to prepare a probe solution with a concentration of 1 mM, and then 1 mM tetrabutylammonium hydroxide was added to obtain the detection reagent.

[0063] A standard solution of 30% hydrogen peroxide was diluted with water to obtain aqueous solutions of hydrogen peroxide at concentrations of 0 mM, 0.2 mM, 0.4 mM, 0.6 mM, 0.8 mM, 1.0 mM, 1.4 mM, 1.6 mM, 1.8 mM, 2.0 mM, 2.5 mM, 3.0 mM, 4.0 mM, and 5.0 mM.

[0064] Measure 1.9 mL of probe detection reagent into a quartz cuvette, then add hydrogen peroxide to dilute it in deionized water to prepare 100 μL of hydrogen peroxide standard aqueous solutions of various concentrations. Take pictures of the solutions with a digital camera. By comparing the collected images, it can be seen that as the concentration of hydrogen peroxide increases, the color of the solution gradually changes from light yellow to gold.

[0065] Example 7

[0066] The probe prepared in step 1 was dissolved in ethanol solvent to prepare a probe solution with a concentration of 1 mM, and then 1 mM tetrabutylammonium hydroxide was added to obtain the detection reagent.

[0067] A standard solution of 30% hydrogen peroxide was diluted with water to obtain aqueous solutions of hydrogen peroxide at concentrations of 0 mM, 0.2 mM, 0.4 mM, 0.6 mM, 0.8 mM, 1.0 mM, 1.4 mM, 1.6 mM, 1.8 mM, 2.0 mM, 2.5 mM, 3.0 mM, 4.0 mM, and 5.0 mM.

[0068] Measure 1.9 mL of probe detection reagent into a quartz cuvette, then add hydrogen peroxide to dilute it in deionized water to prepare 100 μL of hydrogen peroxide standard aqueous solutions of various concentrations. Take pictures of the solutions with a digital camera. By comparing the collected images, it can be seen that as the concentration of hydrogen peroxide increases, the solution gradually changes from green fluorescence to yellow-green fluorescence.

[0069] The above embodiments describe the present invention. It should be understood that the proportions of reagent components in the present invention can be adjusted without departing from the spirit of the present invention. The present invention is also applicable to hydrogen peroxide testing in other fields, and these modifications are also within the scope of the present invention.

Claims

1. The application of a probe for rapid, highly sensitive colorimetric and fluorescence dual-mode detection of hydrogen peroxide, characterized in that, Follow these steps: a. Dissolve the probe N-hexanoic acid-4-borate pinacol ester-1,8-naphthalimide in ethanol solvent to prepare a probe solution with a concentration of 1 mM, and then add 1 mM tetrabutylammonium hydroxide to obtain the detection reagent. b. After adding hydrogen peroxide to the test reagent obtained in step a, the solution color rapidly changes from light yellow to gold within 5 seconds. Under 468nm blue light excitation, the fluorescence changes from green to yellow-green. When the hydrogen peroxide concentration is 0-350 μM, the absorption peak intensity at 454 nm is linearly related to the hydrogen peroxide concentration, and the detection limit is 236.84 nM; when the hydrogen peroxide concentration is 0-80 μM, the fluorescence emission peak intensity at 558 nm is linearly related to the hydrogen peroxide concentration, and the detection limit is 3.36 nM.

Images