N-bromosuccinimide (NBS) detection method based on specific fluorescent probe and application
By using phenothiazine-tetraphenylethylene conjugates as fluorescent probes, the problems of expensive equipment and complex operation of NBS detection are solved, and high-selectivity and low-detection-limit fluorescence detection are achieved, which is suitable for environmental monitoring.
Patent Information
- Application Number
- CN202510950530.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-07-10
- Publication Date
- 2025-10-17
AI Technical Summary
In existing technologies, the detection of N-bromosuccinimide (NBS) mainly relies on high-performance liquid chromatography (HPLC) or mass spectrometry (MS), which has the problems of expensive equipment, complex operation, and difficulty in real-time monitoring, limiting its application in industrial sites or biological systems.
A fluorescent probe with a known compound structure, specifically a phenothiazine-tetraphenylethylene conjugate, was used for fluorescence detection of NBS. The aggregation-induced emission effect in a mixed solvent of water and acetonitrile was utilized, and the fluorescence emission intensity was detected by 365 nm ultraviolet irradiation.
Highly selective fluorescence detection of NBS is achieved with a low detection limit, providing an effective tool for evaluating NBS in environmental monitoring and overcoming the shortcomings of existing technologies such as expensive equipment and complex operation.
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Figure CN120801263A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the field of analytical chemistry and fluorescence sensing technology, and specifically belongs to a fluorescence probe detection method and process for N-bromosuccinimide. BACKGROUND
[0002] N-bromosuccinimide (NBS) is an important brominating reagent and mild oxidant, widely used in organic synthesis, pharmaceutical chemistry and material science. Bromination reaction has the advantages of mild reaction conditions, good selectivity, high yield and easy separation of products, and is applied to free radical bromination of allyl and benzyl, electrophilic bromination of ketones, aromatic compounds or heterocyclic compounds, and addition reaction of olefins. It is also widely used as a catalyst, oxidant, etc. It is a commonly used brominating reagent in chemical and pharmaceutical industries. However, the high reactivity of NBS also brings potential problems, such as: the residues of NBS and its by-products (such as succinimide and bromide) may pollute the environment or affect the safety of drugs; and in pharmaceutical or chemical production, the concentration of NBS needs to be monitored in real time to ensure the reaction efficiency and avoid excessive bromination. At present, the detection of NBS mainly relies on high performance liquid chromatography (HPLC) or mass spectrometry (MS), but these methods have the disadvantages of expensive equipment, complex operation and difficulty in real-time monitoring, which limits their application in industrial sites or biological systems. In recent years, fluorescence probes have become a research hotspot in analytical chemistry due to their high sensitivity, high selectivity, real-time imaging capability and low cost. Therefore, it is of great significance to develop a specific fluorescence probe-based N-bromosuccinimide (NBS) detection method. SUMMARY
[0003] The purpose of the present application is to provide a specific fluorescence probe-based N-bromosuccinimide (NBS) detection method and application of known compound structure, which overcomes the shortcomings of the prior art.
[0004] To solve the above problems, the technical solutions adopted by the present application are as follows: First, a known compound structure as a fluorescence probe in the detection of N-bromosuccinimide, its structural formula is: , The fluorescence probe is a phenothiazine-tetraphenylstilbene conjugated compound, and the molecular formula is C 38 H 27 NS In addition, the fluorescent probe of the present application has an aggregation-induced emission effect in a mixed solvent of water and acetonitrile, and can be applied to the fluorescent detection of NBS; the fluorescent detection step of NBS is: dissolving the fluorescent probe in a solvent to form a fluorescent solution, adding an NBS solution to the above fluorescent solution, then using ultraviolet light with a wavelength of 365 nm for irradiation, and detecting the fluorescence emission intensity.
[0005] The solvent is a mixed solution of water and acetonitrile with a volume ratio of 0-99:100-1.
[0006] Compared with the prior art, the implementation effects of the present application are as follows: the structure of the fluorescent probe in the present application has been reported as a polymer monomer to study the aggregation-induced emission property of the polymer, but its reactivity with NBS has not been studied. In addition, in the prior art, the detection of NBS mainly relies on high performance liquid chromatography (HPLC) or mass spectrometry (MS), and these methods have the disadvantages of expensive equipment and complex operation. The fluorescent material probe of the present application can realize the fluorescent detection of NBS, has good selectivity, and has a low detection limit. The On-Off type fluorescent detection shows that the above-mentioned fluorescent solution can effectively react with different levels of NBS, highlighting its potential as a sensor for detecting NBS, and providing an effective tool for evaluating NBS in environmental monitoring. BRIEF DESCRIPTION OF DRAWINGS
[0007] Figure 1 It is a nuclear magnetic hydrogen spectrum of the fluorescent probe in Example 1 of the present application; Figure 2 It is a nuclear magnetic carbon spectrum of the fluorescent probe in Example 1 of the present application; Figure 3 It is a fluorescence emission spectrum of the fluorescent probe in Example 1 of the present application under 365 nm excitation in a mixed solvent of acetonitrile and water with different proportions; Figure 4 It is a fluorescence emission spectrum of the acetonitrile and water mixed solution of the fluorescent probe in Example 1 of the present application under 365 nm excitation in the presence of different oxides; Figure 5 It is a fluorescence emission spectrum of the acetonitrile and water mixed solution of the fluorescent probe in Example 1 of the present application under 365 nm excitation in the presence of different concentrations of NBS; Figure 6 It is a linear relationship graph of the fluorescence emission peak of the acetonitrile and water mixed solution of the fluorescent probe in Example 1 of the present application under 365 nm excitation in the presence of different concentrations of NBS and the concentration of NBS. DETAILED DESCRIPTION
[0008] With reference to the accompanying drawings on the basis of the embodiments of the present application, the technical solutions in the embodiments of the present application will be described clearly and completely. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments of the present application, all the other embodiments obtained by a person of ordinary skill in the art without creative work fall within the protection scope of the present application. Embodiment one
[0009] Pd2(dba)3, P(O-t-Bu)3 was dissolved in toluene and stirred at room temperature for 10 minutes; the prepared catalyst was added to a toluene solution of phenoxazine, 4-bromotetraphenyl ethylene and t-BuONa, heated to 90°C, and reacted for 12 hours; after the reaction, the reaction solution was cooled to room temperature, extracted with chloroform / water, and the organic phase was rotary evaporated to obtain a bright green fluorescent probe. t t The reaction steps of phenoxazine and 4-bromotetraphenyl ethylene are as follows:
[0010] The reaction steps of phenoxazine and 4-bromotetraphenyl ethylene are as follows:
[0011] The fluorescent probe prepared in this embodiment was analyzed by nuclear magnetic resonance, and its nuclear magnetic resonance hydrogen spectrum (using a Bruker-500MHz nuclear magnetic resonance spectrometer of Bruker Company) was obtained as shown in Figure 1 1 H NMR (500 MHz, CDCl3)δ 7.28 – 7.28 (m, 2H), 7.20 – 7.08 (m, 17H), 7.00 (dd, J = 7.5, 1.7 Hz, 2H), 6.88 (td, J = 7.8, 1.7 Hz, 2H), 6.82 (td, J = 7.4, 1.3 Hz, 2H), 6.16 (dd, J = 8.1, 1.3 Hz, 2H). The fluorescent probe prepared in this embodiment was analyzed by nuclear magnetic resonance, and its nuclear magnetic resonance carbon spectrum (using a Bruker-500MHz nuclear magnetic resonance spectrometer of Bruker Company) was obtained as shown in Figure 2 13 C NMR (125 MHz, CDCI3) δ 144.1, 144.0, 143.6, 143.2, 142.9, 142.1, 140.2, 138.9, 133.6, 131.4, 131.3, 131.3, 130.0, 127.9, 127.8, 127.6, 126.8, 126.7, 126.6, 122.4, 120.1, 117.8, 116.0. The fluorescence emission test and NBS detection test (the test uses a F-4600 fluorescence spectrophotometer of HITACHI) were then performed on the fluorescent probe of the present embodiment: (1) Fluorescence emission of the fluorescent probe in different volume ratios of acetonitrile and water mixed solvents: 5.3 mg of the fluorescent probe was dissolved in 1 mL of acetonitrile, and 10 microliters of the solution was diluted in 10 mL of mixed solvents of acetonitrile and water in different volume ratios, so as to configure a fluorescent probe solution with a concentration of 1 x 10 -5 mol / L. The fluorescence emission of the fluorescent material in different volume ratios of acetonitrile and water mixed solvents was tested under excitation at a wavelength of 365 nm (the volume ratio of water to acetonitrile was 0-99:100-1). The emitted fluorescence was green, and the maximum emission wavelength was 497 nm. The green fluorescence emission spectrum is shown in Figure 3 It can be seen that the fluorescent probe can produce fluorescence emission after being irradiated by ultraviolet light at a wavelength of 365 nm in acetonitrile and water mixed solvents, and the fluorescence intensity is relatively obvious when the water content is 60%-99%.
[0012] (2) Selective detection of NBS by the fluorescent probe A fluorescent probe acetonitrile and water mixed solution (water content 70%) with a concentration of 1 x 10 -5 mol / L was configured. 5 mL of the fluorescent probe solution was taken, and different oxidants (such as HNO3, TEMPO, TBHP, NCS, KMnO4, Fe(NO3)3, Br2, NBS) were added to the above-mentioned fluorescent solution, wherein the concentration of the oxidant was 5 x 10 −5 mol / L, and the fluorescence change was observed. Only in the presence of NBS, the fluorescent solution was quenched under ultraviolet irradiation at a wavelength of 365 nm. The fluorescence emission spectrum is shown in Figure 4 .
[0013] (4) Detection limit determination of the detection of NBS by the fluorescent material 5 mL of a fluorescent probe acetonitrile and water mixed solution (water content 70%) with a concentration of 1 x 10 -5 mol / L was taken, and different concentrations of NBS were added to the above-mentioned fluorescent solution, wherein the concentration of NBS was 0 5 x 10 −5 mol / L, and after irradiation by 365 nm UV light, the change in fluorescence at 497 nm was observed. The fluorescence emission spectrum is shown in Figure 2. Figure 5 The detection limit of the fluorescent solution for NBS was calculated according to the linear relationship between fluorescence intensity and concentration. The linear relationship graph is shown in Figure 3. Figure 6 It can be seen that the fluorescence intensity is linearly related to the concentration of NBS. Example Two
[0014] The difference from Example 1 is that NBS was added to a mixed solution of the fluorescent probe acetonitrile and water (water content 60%) with a concentration of 1 x 10 -5 mol / L, and the change in fluorescence was detected. Example Three
[0015] The difference from Example 1 is that NBS was added to a mixed solution of the fluorescent probe acetonitrile and water (water content 80%) with a concentration of 1 x 10 -5 mol / L, and the change in fluorescence was detected. Example Four
[0016] The difference from Example 1 is that NBS was added to a mixed solution of the fluorescent probe acetonitrile and water (water content 90%) with a concentration of 1 x 10 -5 mol / L, and the change in fluorescence was detected.
[0017] Although embodiments of the present application have been shown and described, it is to be understood that various modifications, substitutions, replacements and variations can be made to these embodiments without departing from the principles and spirit of the present application, and the scope of the present application is defined by the appended claims and their equivalents.
Claims
1. A phenothiazine-tetraphenylethylene conjugate compound is used as a fluorescent probe in the detection of N-bromosuccinimide, characterized in that: The compound structure is: ； The compound is composed of a phenothiazine unit and a tetraphenylethylene unit directly connected by a carbon-nitrogen single bond, and the molecular formula is C 38 H 27 NS.
2. The use according to claim 1, characterized in that The following steps are involved: S1, dissolving the phenothiazine-tetraphenylethylene conjugate compound in a mixed solvent of acetonitrile and water to prepare a fluorescent probe solution with a concentration of 10 μM; S2, adding NBS solutions of different concentrations into the above fluorescent solution; S3, using a UV light source with a wavelength of 365 nm to excite, and detecting the change in fluorescence emission intensity at 497 nm.
3. The detection method according to claim 2, wherein: The ratio of the mixed solvent of acetonitrile and water in the detection system is 0-99:100-1, and the water content is preferably 70%.
4. The detection method according to claim 2, wherein: The concentration of NBS in the detection system was 2-50 μM.
5. The detection method according to claim 2, wherein: The linear detection range of the NBS includes a first linear interval of 2-16 μM and a second linear interval of 18-50 μM, and the detection limit is lower than 10 μM.
6. The use according to claim 2, characterized in that: The fluorescent probe has a specific response to NBS and almost no response to other commonly used oxides (including HNO3, TEMPO, TBHP, NCS, KMnO4, Fe(NO3)3, and Br2) at equimolar concentrations.