A method for detecting Ag based on phenothiazine + and ClO - Fluorescent probe Tz3 and its preparation method and application

Abstract

A method for detecting Ag based on phenothiazine + and ClO ‑ The fluorescent probe Tz3 and its preparation method and application relate to the technical field of fluorescent probes. Sodium carbonate is first added to dichloromethane, and then a mixture of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide and dimethylpyridinamine is added, followed by stirring at room temperature for reaction. The reaction is completed by TLC detection to obtain the fluorescent probe Tz3. The present invention can distinguish Ag + and Hg 2+ , with good selectivity, anti-interference and sensitivity.

Description

Technical Field

[0001] The present invention relates to the technical field of fluorescent probes, and in particular to a method for detecting Ag based on phenothiazine. + and ClO - Fluorescent probe Tz3 and its preparation method and application. Background Art

[0002] Silver (Ag) has been widely used in all aspects of human life. Due to its physical, chemical, and biological advantages, it is widely used in various fields such as photography, agriculture, electronic imaging, medical devices, and the pharmaceutical industry. However, with the widespread use of nanomaterials, silver may enter the soil, water, air, and food, and continue to accumulate in the human body through the food chain. Ag that enters the human body can coordinate with thiol, imidazole, amino, and carboxyl groups, leading to protein and enzyme inactivation, causing many serious symptoms and diseases, and further leading to developmental delay, skin damage, liver and kidney poisoning, and central nervous system damage. Therefore, the development of a silver ion recognition and detection technology has become more important.

[0003] At present, the identification and detection methods of silver ions are mostly ion chromatography and electrochemical methods, such as voltammetry, ICP-MS (inductively coupled plasma mass spectrometry), and AAS (atomic absorption spectroscopy). The above methods are expensive and time-consuming. Therefore, the fluorescent probe detection method has attracted people's attention due to its advantages of simple and fast detection and short response time. However, mercury ions (Hg 2+ ) and silver ions (Ag + ) have similar outer electron configurations. Therefore, when the existing fluorescent probes for detecting silver ions are coordinated with N or O, mercury ions (Hg 2+ ) and silver ions (Ag + ) will respond in the same way; meanwhile, mercury ions (Hg 2+ ) and silver ions (Ag + ) have a high affinity for sulfur, the fluorescent probe designed based on the sulfur-containing receptor will also obtain the same reaction products and signal output when it reacts with mercury ions and silver ions. Therefore, when detecting silver ions, mercury ions will also respond and interfere with the detection results of silver ions. Summary of the Invention

[0004] In order to solve the problem that mercury ions simultaneously respond and interfere with the detection results during the detection of silver ions by fluorescent probes in the prior art, the present invention provides a method for detecting Ag based on phenothiazine. + and ClO - The fluorescent probe Tz3 and its preparation method and application can distinguish Ag + and Hg 2+ , with good selectivity, anti-interference and sensitivity.

[0005] In order to achieve the above-mentioned purpose, the specific scheme adopted by the present invention is: a method for detecting Ag based on phenothiazine + and ClO - The fluorescent probe Tz3 has the structural formula:

[0006] .

[0007] A method for detecting Ag based on phenothiazine + and ClO - The invention discloses a method for preparing the fluorescent probe Tz3. The method is used to prepare the above-mentioned fluorescent probe Tz3. Sodium carbonate is first added to dichloromethane, and then a mixture of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide and dimethylpyridinamine is added, followed by stirring at room temperature for reaction. The reaction is completed by TLC detection to obtain the fluorescent probe Tz3.

[0008] As a kind of phenothiazine-based Ag detection + and ClO - An optimized scheme for the preparation method of the fluorescent probe Tz3: the molar ratio of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide and dimethylpyridinamine in the mixture is 1:1.

[0009] As a kind of phenothiazine-based Ag detection + and ClO - Another optimized scheme for the preparation method of the fluorescent probe Tz3: the ratio of the added amount of the mixture to dichloromethane is 2 mmol:10 ml.

[0010] As a kind of phenothiazine-based Ag detection + and ClO - Another optimized scheme for the preparation method of the fluorescent probe Tz3: the ratio of the added amount of sodium carbonate and dichloromethane is 1 mmol:10 ml.

[0011] A fluorescent probe Tz3 is used to detect Ag + Application in.

[0012] As one of the fluorescent probes mentioned above, Tz3 is used to detect Ag + An optimization scheme for the application of the method: add the fluorescent probe Tz3 solution and the solution to be tested into MeOH:PBS solvent and irradiate with 360-370nm UV lamp for 3-5min. If the color of the solution changes from yellow to colorless and transparent, the solution to be tested contains Ag. + If the color of the solution does not change, the solution to be tested does not contain Ag. + .

[0013] As one of the fluorescent probes mentioned above, Tz3 is used to detect Ag + Another optimization scheme for application in : the concentration of the fluorescent probe Tz3 solution is 10 μM.

[0014] A fluorescent probe Tz3 is used to detect ClO - Application in.

[0015] As one of the fluorescent probes mentioned above, Tz3 can be used to detect ClO - An optimization scheme for the application of the method: add the fluorescent probe Tz3 solution and the solution to be tested into the MeOH:PBS solvent. If the color of the solution changes from yellow to colorless and transparent, the solution to be tested contains ClO - If the color of the solution does not change, the solution to be tested does not contain ClO - .

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] 1. The present invention provides a method for detecting Ag based on phenothiazine + and ClO - The fluorescent probe Tz3 has good specificity and can distinguish Ag + and Hg 2+ , has good selectivity, anti-interference and sensitivity, and at the same time, can respond quickly and adapt to a wide pH range;

[0018] 2. In the present invention, the fluorescent probe Tz3 can also detect ClO - It can be used for detection with good selectivity and anti-interference ability. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 is the H NMR spectrum of the fluorescent probe Tz3 ( 1 HNMR) spectrum;

[0020] Figure 2 The fluorescent probe Tz3 (10 μM) + Ag was added in MeOH:PBS (4:6, v:v; 0.1 μM) solvent. + or UV-visible absorption spectra of other ions (100 μM) + UV;

[0021] Figure 3 The fluorescent probe Tz3 (10 μM) + ClO was added in MeOH:PBS (4:6, v:v; 0.1 μM) solvent. - or UV-visible absorption spectra of other ions (100 μM);

[0022] Figure 4The fluorescent probe Tz3 (10 μM) + Ag was added in MeOH:PBS (4:6, v:v; 0.1 μM) solvent. + or other ions (100 μM) + UV fluorescence spectra (excitation wavelength: 378 nm; slit width: 2.5 nm);

[0023] Figure 5 The fluorescent probe Tz3 (10 μM) + ClO was added in MeOH:PBS (4:6, v:v; 0.1 μM) solvent. - or fluorescence spectra of other ions (100 μM) (excitation wavelength: 378 nm; slit width: 2.5 nm);

[0024] Figure 6 The fluorescent probe Tz3 (10 μM) recognized Ag in the presence of various common metal ion salts in a MeOH:H2O=4:6 solvent. + Fluorescence spectra of the competition experiment of (20μM) (excitation wavelength: 378nm; slit width: 5nm), where M is Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2+ Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2+ 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- , ClO - One of the following; and the horizontal coordinates 1-20 and Ag + 、Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2+ Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2 + 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- , ClO- One-to-one correspondence;

[0025] Figure 7 The fluorescent probe Tz3 (10 μM) recognized ClO in the presence of various common metal ion salts in a MeOH:H2O=4:6 solvent. - Fluorescence spectra of the competition experiment of Ag (20 μM) (excitation wavelength: 378 nm; slit width: 5 nm), where M is Ag + 、Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2+ Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2+ 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- 、NO2 - 、HSO3 - 、SO3 2- One of them, and the horizontal coordinate 1-23 is ClO - 、Ag + 、Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2+ Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2+ 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- 、NO2 - 、HSO3 - 、SO3 2- One-to-one correspondence;

[0026] Figure 8 Ag in MeOH:PBS=4:6 solvent + Linear relationship between the concentration and the fluorescence intensity at 500 nm (excitation wavelength: 378 nm; slit width: 2.5 nm);

[0027] Figure 9 The fluorescent probe Tz3 (10 μM) was reacted with Ag in MeOH:PBS=4:6 solvent. + (30 μM) fluorescence spectra changing with UV irradiation time;

[0028] Figure 10 It is the fluorescent probe Tz3 (10μM) + ClO - Fluorescence spectra of 30 μM as a function of reaction time (excitation wavelength: 378 nm; slit width: 2.5 nm);

[0029] Figure 11 Fluorescent probe Tz3 (10 μM), fluorescent probe Tz3 (10 μM) + Ag + (30μM)+UV, fluorescent probe Tz3 (10μM)+ClO - (30 μM) fluorescence intensity with pH changes;

[0030] Figure 12 ClO in MeOH:PBS=4:6 solvent - Linear relationship between the concentration and the fluorescence intensity at 500 nm (excitation wavelength: 378 nm; slit width: 2.5 nm). DETAILED DESCRIPTION

[0031] The technical scheme of the present invention is further elaborated in detail below in conjunction with specific embodiments. The parts not described and disclosed in detail in the following embodiments of the present invention are all understood to be prior art known or should be known to those skilled in the art. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art; unless otherwise specified, the reagents and materials used in the present invention are all commercially available.

[0032] Example 1

[0033] A method for detecting Ag based on phenothiazine + and ClO - The fluorescent probe Tz3 has the structural formula:

[0034] .

[0035] The nuclear magnetic resonance 1H spectrum of the fluorescent probe Tz3 ( 1 HNMR) spectrum is used to confirm the correctness of its structure. Figure 1Shown, H NMR (400MHz, CDCl3) δ in ppm: 8.57-8.56 (m, 1H), 7.75 (s, 1H), 7.68-7.64 (m, 1H), 7.40 (d, J=7.8Hz, 1H), 7.20-7. 15 (m, 2H), 7.14 (s, 1H), 7.11 (dd, J=7.6, 1.4Hz, 1H), 6.99-6.95 (m, 1H), 6.93 (d, J=8.2Hz , 1H), 6.73 (s, 1H), 3.98 (dd, J=14.0, 7.0Hz, 2H), 3.81 (t, J=4.4Hz, 2H), 3.69 (s, 2H), 3. 43 (t, J=4.8Hz, 2H), 2.60 (t, J=5.0Hz, 2H), 2.55 (t, J=4.4Hz, 2H), 1.47 (t, J=6.9Hz, 3H).

[0036] Example 2

[0037] A method for detecting Ag based on phenothiazine + and ClO - The preparation method of the fluorescent probe Tz3 comprises first adding sodium carbonate to dichloromethane, then adding a mixture of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide and dimethylpyridinamine, and stirring the mixture at room temperature until the reaction is complete as determined by TLC, thereby obtaining the fluorescent probe Tz3. The molar ratio of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide to dimethylpyridinamine in the mixture is 1:1; the ratio of the mixture to dichloromethane is 2 mmol:10 ml; and the ratio of sodium carbonate to dichloromethane is 1 mmol:10 ml.

[0038] The reaction equation during the synthesis process is:

[0039] .

[0040] In this example, 2 mmol of sodium carbonate was first added to 20 ml of dichloromethane, and a mixture of 2 mmol of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide and 2 mmol of dimethylpyridinamine was added to the dichloromethane in the presence of sodium carbonate. The mixture was stirred at room temperature for 3-4 hours. After the reaction was completed by TLC detection, column chromatography was performed using a dichloromethane-methanol system as the mobile phase to obtain a light yellow fluorescent probe Tz3.

[0041] In the preparation method of the fluorescent probe Tz3, 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carboxylic acid is reacted with bromine in chloroform solvent to generate 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide. It should be noted that 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carboxylic acid was prepared according to the method disclosed in the literature "A novel benzothiazine-fused coumarin derivative for sensing hypochlorite with high performance" (Dyes And Pigments, Vol. 182(2020)108675). The preparation method of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carboxylic acid is briefly described as follows: 2-methoxyphenothiazine is reacted with iodoethane in DMSO solvent to generate the intermediate 10-ethyl-2-methoxy-10H-phenothiazine, which is then reacted with phosphorus oxychloride in DMF solvent to generate the intermediate 10-ethyl-2-methoxy-10H-phenothiazine-3-carbaldehyde, which is further reacted with trichloroethane in dichloromethane to generate the intermediate 10-ethyl-2-methoxy-10H-phenothiazine-3-carboxaldehyde. Aluminum chloride reacts and hydrolyzes to generate an intermediate 10-ethyl-2-hydroxy-10H-phenothiazine-3-carboxaldehyde, which is then condensed with diethyl malonate in an acetonitrile solvent to generate an intermediate 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carboxylic acid ethyl ester, which is then hydrolyzed in an ethanol solvent under an alkaline environment to generate 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carboxylic acid.

[0042] Example 3

[0043] A fluorescent probe Tz3 is used to detect Ag + The specific detection process is to add a 10μM fluorescent probe Tz3 solution and a test solution to a MeOH:PBS (4:6, v:v; 0.1μM) solvent, with the volume of the fluorescent probe Tz3 solution and the test solution being 1μL each, and the amount of substance being 1:10; and irradiate with a 360-370nm ultraviolet lamp (UV) for 3-5 minutes. If the color of the solution changes from yellow to colorless and transparent, the test solution contains Ag. + If the color of the solution does not change, the solution to be tested does not contain Ag. + It should be noted that the MeOH:PBS (4:6, v:v; 0.1 μM) solvent means that the volume ratio of MeOH and PBS in the MeOH:PBS solvent is 4:6, and the concentration of PBS in the solvent is 0.1 μM.

[0044] Example 4

[0045] A fluorescent probe Tz3 is used to detect ClO - The specific detection process is to add a 10 μM fluorescent probe Tz3 solution and a test solution into a MeOH:PBS (4:6, v:v; 0.1 μM) solvent. If the solution color changes from yellow to colorless and transparent, the test solution contains ClO. - If the color of the solution does not change, the solution to be tested does not contain ClO - It should be noted that the MeOH:PBS (4:6, v:v; 0.1 μM) solvent means that the volume ratio of MeOH and PBS in the MeOH:PBS solvent is 4:6, and the concentration of PBS in the solvent is 0.1 μM.

[0046] Comparative Example

[0047] A 10 μM solution of fluorescent probe Tz3 and a 100 μM solution of Hg were added to a MeOH:PBS (4:6, v:v; 0.1 μM) solvent. 2+ solution, add fluorescent probe Tz3 solution and Hg 2+ The volume of the solution is 1 μL, the amount of substance is 1:10; and 360-370 nm ultraviolet light (UV) is used for 3-5 min. If the color of the solution does not change, then the fluorescent probe Tz3 does not react with Hg 2+ Interaction, no response.

[0048] <Optional Test>

[0049] 1. Selectivity means that among many ions, the fluorescent probe Tz3 can eliminate interference and only bind to the target ion. A 10 μM fluorescent probe Tz3 solution and a 100 μM test solution were added to a MeOH:PBS (4:6, v:v; 0.1 μM) solvent. The test solution was Ag. + Solution, Al 3+ Solution, Ba 2+ solution, Ca 2+ Solution, Cd 2+ Solution, Co 2+ solution, Cr 3+ solution, Cu 2+ solution, Hg 2+ Solution, Mg 2+ Solution, Mn 2+ Solution, Ni 2+ Solution, Pb 2+ Solution, Pd 2+ Solution, Zn 2+ solution, H2O2 solution, HSO3- Solution, NO2 - Solution, S 2- solution, SO3 2- solution, HCO3 - Solution, Fe 2+ Solution, Fe 3+ One of the solutions was irradiated with a 365 nm ultraviolet lamp (UV) for 3 minutes to obtain the UV-visible absorption spectrum of the solution with different ions added (e.g. Figure 2 ) and fluorescence spectra (as shown in Figure 4 ). Figure 2 It can be seen that only Ag + After interacting with the fluorescent probe Tz3, its UV-visible absorption spectrum shifts blue, specifically, from 428nm to 378nm, and the intensity increases significantly, and the solution color changes from yellow to colorless and transparent; other ions do not interact with the fluorescent probe Tz3, and the solution color remains unchanged. Figure 4 It can be seen that only Ag + After interacting with the fluorescent probe Tz3, it has obvious fluorescence enhancement, a new peak appears at 500nm, and the fluorescence color changes from rose red to green.

[0050] 2. Add 10 μM fluorescent probe Tz3 solution and 100 μM test solution to MeOH:PBS (4:6, v:v; 0.1 μM) solvent. The test solution is ClO - Solution, Ag + Solution, Al 3+ Solution, Ba 2+ solution, Ca 2+ Solution, Cd 2+ Solution, Co 2+ solution, Cr 3+ solution, Cu 2+ solution, Hg 2+ Solution, Mg 2+ Solution, Mn 2+ Solution, Ni 2+ Solution, Pb 2+ Solution, Pd 2+ Solution, Zn 2+ solution, H2O2 solution, HSO3 - Solution, NO2 - Solution, S 2- solution, SO3 2- solution, HCO3 - Solution, Fe 2+ Solution, NO3 - solution, CH3COO - One of the solutions, to obtain the UV-visible absorption spectrum of the solution to be tested with different ions added, such as Figure 3 As shown by Figure 3 It can be seen that only ClO - After interacting with the fluorescent probe Tz3, its UV-visible absorption spectrum blue-shifted, specifically, from 428nm to 378nm, and the intensity increased significantly, and the solution color changed from yellow to colorless and transparent; other ions did not interact with the fluorescent probe Tz3, and the solution color remained unchanged.

[0051] 3. Add 10 μM fluorescent probe Tz3 solution and 100 μM test solution to MeOH:PBS (4:6, v:v; 0.1 μM) solvent. The test solution is ClO - Solution, Ag + Solution, Al 3+ Solution, Ba 2+ solution, Ca 2+ Solution, Cd 2+ Solution, Co 2+ solution, Cr 3+ solution, Cu 2+ solution, Hg 2+ Solution, Mg 2+ Solution, Mn 2+ Solution, Ni 2+ Solution, Pb 2+ Solution, Pd 2+ Solution, Zn 2+ solution, H2O2 solution, HSO3 - Solution, NO2 - Solution, S 2- solution, SO3 2- solution, HCO3 - Solution, Fe 2+ Solution, NO3 - solution, CH3COO - One of the solutions, obtain the fluorescence spectrum of the solution to be tested with different ions added, such as Figure 5 As shown by Figure 5 It can be seen that only ClO - After interacting with the fluorescent probe Tz3, there is a significant fluorescence enhancement, a new peak appears at 500nm, and the fluorescence color changes from rose red to green.

[0052] In summary, the fluorescent probe Tz3 + and ClO - It has good selectivity under the UV-visible absorption spectrum, and the solution changes color, which can be used as a colorimetric probe; Figure 2 It can be seen that Hg 2+ It has no effect on the fluorescent probe Tz3 and the color of the solution does not change; the fluorescent probe Tz3 has no effect on Ag + and ClO- It has good selectivity under the fluorescence spectrum and its fluorescence color also changes, so it can be used as a fluorescent probe. Figure 4 It can be seen that Hg 2+ It has no effect on the fluorescent probe Tz3, no fluorescence enhancement, and no change in fluorescence color. It can detect Ag under ultraviolet light (UV) + Detect ClO without UV light - .

[0053] <Interference test>

[0054] 1. Add 10μM fluorescent probe Tz3 solution and 20μM Ag solution to MeOH:H2O=4:6 solvent. + The solution was irradiated with 365nm ultraviolet light (UV) for 3 minutes, and the fluorescent probe Tz3 and Ag + The reaction showed high intensity fluorescence value ( Figure 6 The area filled with diagonal lines at the horizontal coordinate 1).

[0055] A 10 μM fluorescent probe Tz3 solution and a 50 μM test solution were added to a MeOH:H2O=4:6 solvent and irradiated at 365 nm UV light for 3 minutes. The test solution was M+Ag. + solution, Figure 6 The oblique line filled areas at the horizontal coordinates 2-20 are the fluorescent probes Tz3 and M+Ag respectively. + The fluorescence value of the reaction; where M is Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2 + Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2+ 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- , ClO - One of the Figure 6 The horizontal coordinate 2-20 and Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2+ Cr 3+ 、Cu2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2+ 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- , ClO - One to one correspondence. Figure 6 It can be seen that in Ag + In the presence of other ions, the fluorescent probe Tz3 still maintains a high intensity fluorescence value ( Figure 6 The area filled with oblique lines at the horizontal coordinates 2-20), where Figure 6 Ag at the horizontal coordinate 19 + and S 2- Chemical precipitation occurs, resulting in the fluorescent probe Tz3 and S 2- +Ag + The fluorescence value of the reaction is small.

[0056] A 10 μM fluorescent probe Tz3 solution and a 50 μM test solution were added to a MeOH:H2O=4:6 solvent and irradiated with a 365 nm ultraviolet lamp (UV) for 3 minutes. The test solution contained only M, and M was Al. 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2 + Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2+ 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- , ClO - One of Figure 6 The solid filled areas at 2-20 in the middle horizontal axis are the fluorescence values ​​of the fluorescent probe Tz3 when it reacts with M; the fluorescent probe Tz3 has almost no fluorescence value when it reacts with other ions (such as Figure 6 The solid filled area at the horizontal axis 2-20 in the middle) indicates that the fluorescent probe Tz3 has strong selectivity and anti-interference to silver ions, and under ultraviolet light (UV) irradiation, the fluorescent probe Tz3 has strong selectivity and anti-interference to ClO - No response.

[0057] 2. Add 10 μM fluorescent probe Tz3 and 20 μM ClO to MeOH:H2O=4:6 solvent. - Solution, fluorescent probe Tz3 and ClO - The reaction showed high intensity fluorescence value ( Figure 7 The area filled with diagonal lines at the horizontal coordinate 1).

[0058] The fluorescent probe Tz3 with a concentration of 10 μM and the detection solution with a concentration of 50 μM were added to the MeOH:H2O=4:6 solvent. The detection solution was M+ClO - Solution, that is, the solution to be tested contains M and ClO - , Figure 7 The oblique line filled areas at the horizontal coordinates 2-23 are the fluorescent probes Tz3 and M+ClO - The fluorescence value of the reaction, where M is Ag + 、Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2+ Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2+ 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- 、NO2 - 、HSO3 - 、SO3 2- One of Figure 7 The horizontal coordinate 2-23 and Ag + 、Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2+ Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2 + 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S2- 、NO2 - 、HSO3 - 、SO3 2- One to one correspondence. Figure 7 It can be seen that in ClO - In the presence of other ions, the fluorescent probe Tz3 still maintains a high intensity fluorescence value ( Figure 7 The area filled with diagonal lines at the horizontal coordinate 2-23).

[0059] Add 10μM fluorescent probe Tz3 and 50μM test solution to MeOH:H2O=4:6 solvent. The test solution contains only M, and M is Ag. + 、Al 3+ 、Ba 2+ , Ca 2+ 、Cd 2+ 、Co 2+ Cr 3+ 、Cu 2+ 、Fe 2+ 、Fe 3+ 、Hg 2+ Mg 2+ 、Mn 2+ 、Ni 2+ , Pb 2+ 、Pd 2+ 、Zn 2+ 、H2O2、S 2- 、NO2 - 、HSO3 - 、SO3 2- One of Figure 7 The solid filled areas at the horizontal coordinates 2-23 are the fluorescence values ​​of the fluorescent probe Tz3 reacting with M. Figure 7 It can be seen that the fluorescent probe Tz3 has almost no fluorescence value when reacting with other ions (such as Figure 7 The solid filled area at the horizontal coordinate 2-23 in the middle) and in the absence of ultraviolet (UV) irradiation, the fluorescence probe Tz3 + No response, indicating that the fluorescent probe Tz3 is sensitive to ClO - It has strong selectivity and anti-interference ability.

[0060] In summary, the fluorescent probe Tz3 is sensitive to silver ions and ClO - It has good selectivity and anti-interference under fluorescence spectrum. Figure 6The area filled with diagonal lines at the horizontal axis 1 represents the detection result of fluorescent probe Tz3 on silver ions, the area filled with diagonal lines at the horizontal axis 11 represents the detection result of fluorescent probe Tz3 on the mixture of silver ions and mercury ions, and the solid filled column at the horizontal axis 11 represents the detection result of fluorescent probe Tz3 on mercury ions. The results show that when fluorescent probe Tz3 detects silver ions, mercury ions do not respond and will not affect the detection results. - It has good selectivity and anti-interference in fluorescence spectrum. At the same time, the fluorescent probe Tz3 has good selectivity and anti-interference in Ag under ultraviolet light. + There is a response to ClO - No response; In the absence of UV light, the fluorescent probe Tz3 reacts with ClO - There is a response to Ag + No response; that is, the fluorescent probe Tz3 can well distinguish Ag with or without UV light irradiation. + and ClO - .

[0061] <Detection Limit Test>

[0062] Preparation of different concentrations of Ag + Fluorescent probe Tz3 solution and different concentrations of Ag were added to MeOH:PBS=4:6 solvent. + The solution was irradiated with 365nm ultraviolet light (UV) for 3 minutes to obtain different Ag + The linear relationship between the solution and the fluorescence intensity at 500 nm (such as Figure 8 As shown), the linear relationship between the fluorescent probe Tz3 and Ag is obtained. + The fluorescence detection limit concentration was 1.87 nM.

[0063] Preparation of different concentrations of ClO - Fluorescent probe Tz3 solution and different concentrations of ClO were added to MeOH:PBS=4:6 solvent. - solution, and obtain different ClO - The linear relationship between the solution and the fluorescence intensity at 500 nm (such as Figure 12 As shown in the figure), the linear relationship between the fluorescent probe Tz3 and ClO - The fluorescence detection limit concentration is 0.85 nM.

[0064] In summary, the fluorescent probe Tz3 + and ClO - The detection limit is very low, enough to detect trace amounts of Ag + and ClO - .

[0065] <Response time test>

[0066] Add a 10 μM fluorescent probe Tz3 solution to a MeOH:PBS = 4:6 solvent and irradiate it with a 360 - 370 nm ultraviolet lamp (UV) for different times to obtain fluorescence spectra that change with the irradiation time of the ultraviolet lamp (UV) (as Figure 9 shown); add a 10 μM fluorescent probe Tz3 solution and a 30 μM Ag + solution to a MeOH:PBS = 4:6 solvent, and the molar ratios of Ag + to the fluorescent probe Tz3 are 1:1, 3:1, and 5:1 respectively. Then irradiate it with a 360 - 370 nm ultraviolet lamp (UV) for different times to obtain fluorescence spectra that change with the irradiation time of the ultraviolet lamp (UV) (corresponding to Figure 9 1eqAg + 、3eqAg + 、5eqAg + in

[0067] Add a 10 μM fluorescent probe Tz3 solution to a MeOH:PBS = 4:6 solvent and react for different times to obtain fluorescence spectra that change with the reaction time (as Figure 10 shown); add a 10 μM fluorescent probe Tz3 solution and a 30 μM ClO - solution to a MeOH:PBS = 4:6 solvent and react for different times. Among them, the mass ratios of ClO - to the fluorescent probe Tz3 are 1:1, 3:1, and 5:1 respectively, to obtain fluorescence spectra that change with the reaction time (corresponding to Figure 10 1eqClO - 、3eqClO - 、5eqClO - in

[0068] The reaction time of the fluorescent probe Tz3 with Ag + reaches the maximum fluorescence intensity at 4 min, and the reaction time of the fluorescent probe Tz3 with ClO - reaches the maximum fluorescence intensity at 30 s. The response time of the fluorescent probe Tz3 to both is very short, and it can complete the detection within a short time.

[0069] <pH Influence Test>

[0070] Under different pH conditions, use the fluorescent probe Tz3 to detect Ag + and ClO - , and the obtained results (as Figure 11 shown), as shown by Figure 11 , the fluorescent probe Tz3 can maintain a good response at pH = 4 - 10. Therefore, the fluorescent probe Tz3 can achieve the detection of Ag under amphoteric acid-base conditions.+ and ClO - Detection of two ions.

[0071] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for detecting Ag based on phenothiazine + and ClO - The fluorescent probe Tz3 is characterized by: The structural formula of the fluorescent probe Tz3 is:

2. A method for detecting Ag based on phenothiazine + and ClO - A method for preparing the fluorescent probe Tz3 according to claim 1, wherein: Sodium carbonate was first added to dichloromethane, and then a mixture of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide and dimethylpyridinamine was added, followed by stirring at room temperature. The reaction was completed as detected by TLC to obtain the fluorescent probe Tz3; the molar ratio of 11-ethyl-2-oxo-2,11-dihydropyrano[2,3-b]phenothiazine-3-carbonyl bromide to dimethylpyridinamine in the mixture was 1:

1.

3. A method for detecting Ag based on phenothiazine as claimed in claim 2 + and ClO - The preparation method of the fluorescent probe Tz3 is characterized by: The ratio of the added amount of the mixture to that of dichloromethane was 2 mmol:10 ml.

4. A method for detecting Ag based on phenothiazine as claimed in claim 2 + and ClO - The preparation method of the fluorescent probe Tz3 is characterized by: The ratio of sodium carbonate to dichloromethane added is 1mmol:10ml.

5. A fluorescent probe Tz3 as claimed in claim 1 for detecting Ag + The application is characterized by: Add the fluorescent probe Tz3 solution and the solution to be tested into MeOH:PBS solvent and irradiate with 360-370nm UV lamp for 3-5min. If the color of the solution changes from yellow to colorless and transparent, the solution to be tested contains Ag. + If the color of the solution does not change, the solution to be tested does not contain Ag. + .

6. The fluorescent probe Tz3 as claimed in claim 5 is used to detect Ag + The application is characterized by: The concentration of the fluorescent probe Tz3 solution was 10 μM.

7. A fluorescent probe Tz3 as claimed in claim 1 for detecting ClO - The application is characterized by: Add the fluorescent probe Tz3 solution and the test solution into the MeOH:PBS solvent. If the solution changes from yellow to colorless and transparent, the test solution contains ClO. - If the color of the solution does not change, the solution to be tested does not contain ClO - .

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