CN112939956A - Fluorescent probe for detecting mercury ions and hypochlorite ions and preparation method and application thereof - Google Patents
Fluorescent probe for detecting mercury ions and hypochlorite ions and preparation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of organic synthesis, and particularly relates to a fluorescent probe for detecting mercury ions and hypochlorite ions, and a preparation method and application thereof. The molecular formula of the fluorescent probe is C19H19NO3S2The structural formula is as follows:
Description
Technical Field
The invention belongs to the technical field of organic synthesis, and particularly relates to a fluorescent probe for detecting mercury ions and hypochlorite ions, and a preparation method and application thereof.
Background
Mercury compounds can react with nucleic acids, enzymes and proteins in the organism, causing damage to the organism. Hypochlorous acid itself has strong nucleophilicity and oxidation, and plays an important role in immune defense of a living body. Therefore, the method has very important significance in simply, reliably and accurately detecting the mercury ions and the hypochlorite ions.
The existing methods for measuring the content of mercury and hypochlorous acid are various, such as a colorimetric method, an iodometry method, an enzyme circulation method, a capillary electrophoresis method, an HPLC method and the like, and the methods have advantages but have disadvantages. Colorimetric methods are based on a color reaction that produces a colored compound, and generally include two steps: firstly, selecting a proper chromogenic reagent to react with a component to be measured to form a colored compound, and then comparing or measuring the color depth of the colored compound, wherein in the process, the acidity of a solution, the dosage of the chromogenic reagent, the temperature, a solvent and the like have influence on the color reaction, and the measurement sensitivity and accuracy are poor; the iodometry is a widely applied method in the redox titration method, but is sensitive to the acidity and temperature reaction of a sample solution and is not easy to operate; the enzyme cycling method is a measuring method for amplifying a measured substance by utilizing the substrate specificity of enzyme, but the used tool enzyme has large using amount and higher cost; the capillary electrophoresis method is an electrophoresis separation analysis method which takes an elastic quartz capillary as a separation channel, takes a high-voltage direct-current electric field as a driving force and realizes separation according to the difference of mobility and distribution behavior among components in a sample, the capillary has small diameter, so that the optical path is too short, the sensitivity is low, electroosmosis can be changed due to the composition of the sample, the separation reproducibility is further influenced, and the defects can cause errors of disease diagnosis; the HPLC method has high accuracy, simple sample processing, limitation of chromatographic column and long time consumption.
While the fluorescent probe detection method is valued by researchers with the advantages of nondestructive and visual in-situ detection, the fluorescent probe which can specifically respond to mercury ions and hypochlorite ions and can distinguish the mercury ions from the hypochlorite ions has not been reported.
Disclosure of Invention
In order to overcome the problems in the prior art, the invention provides a fluorescent probe for detecting mercury ions and hypochlorite ions. The fluorescent probe realizes specific recognition response to mercury ions and hypochlorite ions, is not influenced by interference ions, and can judge whether the mercury ions and the hypochlorite ions exist or not by direct visual observation.
The invention also provides a preparation method and application of the fluorescent probe.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a fluorescent probe for detecting mercury ions and hypochlorite ions, wherein the molecular formula of the fluorescent probe is C19H19NO3S2The structural formula is as follows:
the above synthetic route for detecting mercury ions and hypochlorite ions is as follows:
the preparation method specifically comprises the following steps:
(1) adding 4-bromo-1, 8-naphthalic anhydride and n-butylamine into ethanol, refluxing under the protection of nitrogen until the reaction solution is clear, concentrating, cooling to separate out crystals, and recrystallizing to obtain a compound 1;
(2) dissolving the compound 1 prepared in the step (1) in methanol, adding sodium methoxide and copper sulfate, performing reflux reaction for 11-15 h, concentrating, cooling, precipitating crystals, and washing with deionized water to obtain a compound 2;
(3) adding the compound 2 prepared in the step (2) into HI, performing reflux reaction for 12-24 h, cooling, filtering and washing to obtain a compound 3;
(4) adding the compound 3 prepared in the step (3), urotropine and paraformaldehyde into acetic acid, carrying out reflux reaction for 1-3 h, cooling to 80-90 ℃, adding excessive concentrated hydrochloric acid, continuing to react for 35-40 min, cooling, filtering to obtain filtrate, extracting with dichloromethane to obtain a crude product, and separating the crude product by using column chromatography to obtain a compound 4;
(5) adding the compound 4 prepared in the step (4), anhydrous magnesium sulfate and p-toluenesulfonic acid into tetrahydrofuran, and stirring under the protection of nitrogen to obtain a mixed solution; dissolving 1, 4-ethanedithiol in tetrahydrofuran, adding the tetrahydrofuran into the mixed solution, stirring at normal temperature for 6-8 h, removing the solvent after the reaction is finished, and performing column chromatography separation to obtain the fluorescent probe.
Preferably, the molar ratio of the 4-bromo-1, 8-naphthalic anhydride to the n-butylamine in the step (1) is 1 (5-7).
Preferably, the molar ratio of the compound 1 to the sodium methoxide in the step (2) is 1 (8-12).
Preferably, the amount of the compound 2 in the step (3) is 1.5-2.0 g, and the amount of HI is 20-40 mL.
Preferably, the molar ratio of the compound 3 to the urotropine in the step (4) is 1 (1-1.5); the mass ratio of the urotropine to the paraformaldehyde is 1 (1-1.5).
Preferably, the molar ratio of the compound 4, the anhydrous magnesium sulfate, the p-toluenesulfonic acid and the 1, 4-ethanedithiol in the step (5) is 1 (1-3): (1-1.2): 1-1.2).
The fluorescent probe is applied to the fluorescent detection of mercury ions and hypochlorite ions, and is particularly used for the fluorescent detection, visual qualitative detection and cell imaging detection of the content of the mercury ions and the hypochlorite ions.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention designs and synthesizes naphthalimide-1, 4-ethyl disulfide fluorescent probe by using novel naphthalimide dye, in a PBS system, the whole molecule of the fluorescent probe is faint yellow and weak yellow fluorescent, along with the continuous increase of the concentration of mercury ions and hypochlorite ions of specific analytes in the system, 1, 4-ethyl disulfide is induced to react with mercury ions and hypochlorite ions, so that 1, 4-ethyl disulfide and mercury ions form insoluble matters, or 1, 4-ethyl disulfide and hypochlorite ions react, the fluorescent probe releases naphthalimide dye molecular monomers, the fluorescence intensity is obviously enhanced and along with obvious color change, and selected interference ions and the like almost have no influence on the detection effect, thereby realizing the specific recognition response to the mercury ions and hypochlorite ions;
2. the invention can judge whether mercury ions and hypochlorite ions exist or not by direct visual observation, and has wide application prospect in the field of biomolecule detection.
Drawings
FIG. 1 is a nuclear magnetic hydrogen spectrum of a fluorescent probe prepared in example 1;
FIG. 2 is a nuclear magnetic carbon spectrum of the fluorescent probe prepared in example 1;
in FIG. 3, (a) is a graph showing the change of the fluorescence spectrum of the fluorescent probe prepared in example 1 and 10 equivalents of mercury ions with the increase of the reaction time, and (b) is a graph showing the change of the fluorescence intensity at 520nm with the increase of the reaction time (Ex 410nm, Em 400-750nm, Ex. slit/Em. slit 2.5/2.5);
fig. 4 (a) is a graph showing the change of the fluorescence spectrum of the fluorescent probe prepared in example 1 after reacting with different equivalents of mercury ions, and (b) is the change of the fluorescence intensity at 520nm with the addition of mercury ions (Ex 410nm, Em 400-750nm, Ex. slit/Em. slit 2.5/2.5);
FIG. 5 shows the results of changes in fluorescence spectra of the fluorescent probe prepared in example 1 after reaction with different cations (Ex 410nm, Em 400-750nm, Ex. slit/Em. slit 2.5/2.5);
FIG. 6 is a bar graph of fluorescence intensity at 520nm after reaction of the fluorescent probe prepared in example 1 with different cations (Ex 410nm, Em 400-750nm, Ex. slit/Em. slit2.5/2.5), in which: from left to right (1-19) are blank control and Hg2+、K+、Na+、Al3+、Mg2+、Fe3+-、Ag+、Cd2+、Co2+、Zn2+、Cs2+、Mn2+、Ca2+、Sr2+、Cu2+、Sn2+、Ba2+、Ni3+；
FIG. 7 is a color change of a solution before and after a reaction of the fluorescent probe prepared in example 1 with mercury ions under a fluorescent lamp; wherein: (a) before the fluorescent probe reacts with mercury ions, and (b) after the fluorescent probe reacts with mercury ions;
FIG. 8 is a color change of a solution under an ultraviolet lamp before and after a reaction of the fluorescent probe prepared in example 1 with mercury ions; wherein: (a) before the fluorescent probe reacts with mercury ions, and (b) after the fluorescent probe reacts with mercury ions;
FIG. 9 is a UV absorption spectrum of a fluorescent Probe Probe prepared in example 1 with time after hypochlorous acid was added;
FIG. 10 shows the change of fluorescence spectrum of the fluorescent probe prepared in example 1 with 0-0.5 equivalent of hypochlorite ion with the increase of reaction time, and (b) shows the change of fluorescence intensity at 560nm with the increase of reaction time (Ex 430nm, Em 440-750nm, Ex. slit/Em. slit 2.5/2.5);
FIG. 11 (a) is a graph showing the change in fluorescence spectrum of the fluorescent probe prepared in example 1 after reacting with 0.5 to 1.5 equivalents of hypochlorite ions; (b) the change of the fluorescence intensity at 530nm along with the addition equivalent of mercury ions (Ex 430nm, Em 440-750nm, Ex. Slit/Em. Slit 2.5/2.5);
FIG. 12 shows the results of changes in fluorescence spectra of the fluorescent probe prepared in example 1 after reaction with different anions (Ex 430nm, Em 440 750nm, Ex. slit/Em. slit 2.5/2.5);
FIG. 13 is a bar graph of fluorescence intensity at 530nm after reaction of the fluorescent probe prepared in example 1 with different anions (Ex 430nm, Em 440-750nm, Ex. slit/Em. slit2.5/2.5), in which: blank control and ClO are sequentially arranged from left to right (1-17)-、F-、Cl-、Br-、I-、CN-、PO4 3-、Ac-、SO4 2-、CO3 2-、HSO4 -、SCN-、HS-、SO3 2-、HCO3 -、NO3 -；
FIG. 14 is a color change of a solution before and after the reaction of the fluorescent probe prepared in example 1 with hypochlorite ions under a fluorescent lamp; wherein: (a) before the fluorescent probe reacts with hypochlorite ions, (b) after the fluorescent probe reacts with hypochlorite ions;
FIG. 15 is a color change of a solution under an ultraviolet lamp before and after a reaction of a fluorescent probe prepared in example 1 with hypochlorite ions; wherein: (a) before the fluorescent probe reacts with hypochlorite ions, and (b) after the fluorescent probe reacts with hypochlorite ions.
Detailed Description
The invention is further illustrated, but not limited, by the following examples and the accompanying drawings.
Example 1
The preparation method of the fluorescent probe comprises the following steps:
(1) respectively weighing 4.10g (15mmol) of 4-bromo-1, 8-naphthalic anhydride and 1.12g (15mmol) of n-butylamine into 30ml of ethanol, carrying out reflux reaction under the protection of nitrogen until reaction liquid is clear, cooling after concentration to separate out crystals, and recrystallizing with ethanol to obtain the compound 1, wherein the yield is 14.41g, the yield is 89%, and the structural formula is as follows:
(2) dissolving 3.31g (10mmol) of the compound 1 prepared in the step (1) in 20ml of methanol, sequentially adding 0.70(10mmol) of sodium methoxide and 0.25g (1mmol) of copper sulfate pentahydrate, refluxing for 12 hours, concentrating, cooling to separate out crystals, and washing with deionized water to obtain a compound 2 with the yield of 2.51g and the yield of 87.2 percent, wherein the structural formula is as follows:
(3) adding 2.13g (7.5mmol) of the compound 2 prepared in the step (2) into 30mL of concentrated HI (57 mass percent), refluxing and reacting at 140 ℃ for 8h (the compound 2 is completely reacted by monitoring by TLC), cooling, filtering and washing with diethyl ether to obtain the compound 3, wherein the yield is 1.38g, the yield is 93%, and the structural formula is as follows:
(4) putting 200.4mg (0.744mmol) of compound 3, 104.3mg (0.744mmol) of urotropine and 104.3mg of paraformaldehyde in a 50mL round-bottom flask, adding excessive acetic acid (about 15 mL), refluxing for 1h, cooling to 85 ℃, adding excessive concentrated hydrochloric acid (about 15mL in mass fraction) and reacting for 35 min; cooling the reaction liquid to room temperature, adding deionized water for dilution, filtering to obtain filtrate, extracting with dichloromethane to obtain a crude product, then separating by using a silica gel column to obtain a compound 4, wherein the size of silica gel particles is 200-300 meshes, the ratio of eluent for column separation is petroleum ether and ethyl acetate is 3:1 (volume ratio, the same below), the yield of the compound 4 is 80.69%, and the structural formula is as follows:
(5) adding a compound 4(300mg, 1.02mmol), anhydrous magnesium sulfate (1g, 8.30mmol) and p-toluenesulfonic acid (210mg, 1.20mmol) into 10ml of tetrahydrofuran, stirring for 10min under the protection of nitrogen to obtain a mixed solution, dissolving 1, 4-ethanedithiol (213.05mg, 1.22mmol) into 5ml of anhydrous tetrahydrofuran, slowly dropwise adding the obtained solution into the mixed solution, stirring for 7h at normal temperature, and completely protecting with nitrogen; and then separating by silica gel column chromatography to obtain the fluorescent probe, wherein the size of the silica gel particle is 200-300 meshes, the eluent petroleum ether and dichloromethane are 1:4, the yield is 59.61%, and the structural formula of the fluorescent probe is as follows:
fig. 1 and 2 are a nuclear magnetic hydrogen spectrum and a nuclear magnetic carbon spectrum of the fluorescent probe prepared in this example, respectively. The results of structural characterization according to fig. 1 and 2, wherein the carbon spectra data are:13C NMR(101MHz,CDCl3) δ 164.35(s),163.74(s),158.22(s),134.14(s),132.00(s),129.47(s),129.00(s),126.17(s),123.79(s),122.40(s),115.13(s),114.60(s),55.87(s),40.20(d, J ═ 11.5Hz),40.14(s),30.27(s),20.40(s), and 13.88(s).
Fluorescence detection application assay
(1) Reagent preparation
a. Fluorescent probe solution (1.00X 10)-3mol·L-1) The preparation of (1):
the fluorescent probe prepared in example 1 was dissolved in ethanol to prepare a solution having a concentration of 1.00X 10-3mol·L-1The fluorescent probe solution of (1);
b. preparation of standard solutions of various cations and anions:
standards of various cations and anions were formulated with deionized water to a concentration of 1.00X 10-3mol·L-1In addition, mercury chloride and sodium hypochlorite are prepared into a part with the concentration of 1.00X 10-2mol·L-1A standard solution of (4);
c. detection of fluorescent probes and Mercury ion Properties
B, taking out 30 mu L of the fluorescent probe solution prepared in the step a, adding the fluorescent probe solution into a cuvette, and adding the fluorescent probe solution with the concentration of 1.00 multiplied by 10-2mol·L-1The standard solution of mercury chloride is diluted to 3mL by ethanol, and the fluorescence property of the standard solution of mercury chloride is measured once every 1min until the fluorescence intensity is not enhanced any more (Ex 410nm, Em 400-750nm, Ex. slit/Em. slit2.5/2.5), and the obtained fluorescence spectrum is as shown in (a) in fig. 3, and as can be seen from (a) in fig. 3, the fluorescence intensity is gradually enhanced along with the increase of the reaction time, which indicates that a certain time effect exists in the action of the probe and mercury ions, and provides reliable action time for other responsiveness studies of the probe. FIG. 3(b) is a graph showing the increase in fluorescence intensity at 520nm with the reaction timeIn the case of change, the fluorescence intensity reaches the maximum value within 30min as time increases, and as shown in FIG. 3(b), the fluorescence intensity does not change any more with time, and the fluorescence intensity increases by 14 times compared with that before the addition of mercury ions, and the maximum emission wavelength undergoes blue shift from 560nm to 530 nm.
Taking 30 groups of 30 mu L of the fluorescent probe solution prepared in the step a, and sequentially adding the 30 groups of 30 test tubes into 1 test tube as a blank control without adding any component, and sequentially adding 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.2, 2.4, 2.6, 2.8, 3.3, 3.6, 4.0, 4.5 and 5 equivalents of the fluorescent probe solution into the rest 35 test tubes to obtain a concentration of 1.00 multiplied by 10-3mol·L-1The fluorescence property of the probe (Ex 410nm, Em 400-750nm, Ex. slit/Em. slit2.5/2.5) was measured by diluting the standard solution with ethanol to 3mL, and the fluorescence spectrum was as shown in fig. 4 (a), the fluorescence intensity gradually increased with the increase of the added mercury ion equivalent, and the fluorescence intensity was in a linear relationship with the mercury ion equivalent as shown in fig. 4 (b), and the detection limit of the probe was found to be 0.04 μ M by calculating the formula D3 σ/k (where σ is the blank standard deviation and k is the slope of the fitting curve) according to the detection limit.
Taking out 19 groups of 30 mu L of the fluorescent probe solution prepared in the step a, adding the 19 groups of 30 mu L of the fluorescent probe solution into 19 test tubes respectively, taking 1 test tube as a blank control, adding no component, and sequentially adding 10 equivalents of the fluorescent probe solution prepared in the step b into the rest 18 test tubes to obtain a solution with the concentration of 1.00 multiplied by 10-3mol·L-1The following standard solutions: hg is a mercury vapor2+；K+；Na+；Al3+；Mg2+；Fe3+；Ag+；Cd2+；Co2+；Zn2+；Cs2+；Mn2+；Ca2+；Sr2+；Cu2+；Sn2+；Ba2+；Ni3+Then, the solution was diluted to 3mL with ethanol, and the fluorescence emission spectrum of the solution was measured after 120min (Ex 410nm, Em 400-750nm, Ex. slit/Em. slit2.5/2.5), with the results shown in fig. 5; FIG. 6 shows a fluorescent probe andhistograms of fluorescence intensity at 520nm after different cation reactions (from left to right (i.e. from numbers 1-19) are blank, Hg2+、K+、Na+、Al3+、Mg2+、Fe3+-、Ag+、Cd2+、Co2+、Zn2+、Cs2+、Mn2+、Ca2 +、Sr2+、Cu2+、Sn2+、Ba2+、Ni3+)。
As can be seen from FIGS. 5 and 6, other ions have almost no fluorescent response to the fluorescent probe described in this embodiment, and the mercury ions and the fluorescent probe have significant fluorescent response, which indicates that the fluorescent probe of the present invention has high selectivity for detection of mercury ions, and can be used for detection of mercury ions.
B, taking out 30 mu L of the fluorescent probe solution prepared in the step a, adding the fluorescent probe solution into a test tube, and adding the fluorescent probe solution into the test tube at a concentration of 1.00 multiplied by 10-2mol·L-1The standard solution of mercury chloride is diluted to 3mL with ethanol, as shown in fig. 7, under a fluorescent lamp, visible to naked eyes, mercury ions can make the ethanol solution of the fluorescent probe undergo a significant color change, and the color of the solution changes from orange (in the figure (a)) to yellow-green (in the figure (b)); as shown in fig. 8, the fluorescent probe emits yellow-green fluorescence under the ultraviolet lamp, which is visible to the naked eye, and the mercury ion induces the fluorescent probe to emit yellow-green fluorescence (b) in the figure, indicating that the fluorescent probe of the present invention is a fluorescent probe with a color generation sensing function, and can be used for visual qualitative detection of mercury ions, thereby greatly facilitating the rapid qualitative detection of mercury ions.
d. Detection of fluorescent probes and hypochlorite ion Properties
B, taking out 30 mu L of the fluorescent probe solution prepared in the step a, adding the fluorescent probe solution into a cuvette, and adding the fluorescent probe solution with the concentration of 1.00 multiplied by 10-3mol·L-1The sodium hypochlorite standard solution (15. mu.L) was diluted to 3mL with ethanol, and then the ultraviolet absorption spectrum was measured every 1min until the absorbance was not increased any more, and the obtained ultraviolet spectrum is shown in FIG. 9, as shown in FIG. 9, the maximum absorption wavelength was blue-shifted from 460nm to 435nm with the increase of the reaction time, and the absorption intensity was slightly decreased. The reaction speed is fast, after the hypochlorous acid is added,the reaction is completed within 10 seconds, the time is prolonged, the absorption spectrum does not change any more, and the probe can rapidly identify hypochlorite ions.
Taking 16 groups of 30 mu L of the fluorescent probe solution prepared in the step a, sequentially adding the 16 groups of 30 mu L of the fluorescent probe solution into 30 test tubes, wherein 1 test tube is used as a blank control, no component is added, and 0 equivalent, 0.1 equivalent, 0.2 equivalent, 0.3 equivalent, 0.4 equivalent, 0.5 equivalent, 0.6 equivalent, 0.7 equivalent, 0.8 equivalent, 0.9 equivalent, 1.0 equivalent, 1.1 equivalent, 1.2 equivalent, 1.3 equivalent, 1.4 equivalent and 1.5 equivalent are sequentially added into the rest 15 test tubes-3mol·L-1The fluorescence properties of the sodium hypochlorite standard solution (Ex 430nm, Em 440-750nm, Ex. slit/Em. slit2.5/2.5) were measured by diluting the solution to 3mL with ethanol, and the fluorescence spectra are respectively shown in fig. 10 (a) and fig. 11 (a), and it can be seen that the fluorescence intensity gradually increases with the increase of the equivalent of hypochlorite ion added, and when 0.5 equivalent is added, the fluorescence intensity reaches the peak, and when hypochlorite ion is continuously added, the fluorescence gradually decreases to 0-5 μ M, and as shown in fig. 10 (b) and fig. 11 (b), the fluorescence intensity and the equivalent of hypochlorite ion have a linear relationship, and the detection limit of the formula D3 σ/k (where σ is the blank standard deviation and k is the slope of the fitting curve) reaches 0.01 μ M.
Taking 18 groups of 30 mu L of the fluorescent probe solution prepared in the step a, adding the 18 groups of 30 mu L of the fluorescent probe solution into 17 test tubes respectively, wherein 1 test tube is used as a blank control, no component is added, and 0.5 equivalent of the fluorescent probe solution is added into 1 test tube with the concentration of 1.00 multiplied by 10- 3mol·L-1The rest 15 test tubes are sequentially added with the 10 equivalent sodium hypochlorite standard solution prepared in the step b, and the concentration of the solution is 1.00 multiplied by 10-3mol·L-1Standard solution F of-；Cl-；Br-；I-；CN-；PO4 3-；Ac-；SO4 2-；CO3 2-；HSO4 -；SCN-；HS-；SO3 2-；HCO3 -；NO3 -Then, the solution was diluted to 3mL with ethanol, and the fluorescence emission spectrum of the solution was measured after 120min (Ex ═ 430nm, Em ═ 440-;FIG. 13 is a bar graph of fluorescence intensity at 530nm after reaction of fluorescent probes with different anions (from left to right (i.e., numbered from 1-17) as blank, ClO-、F-、Cl-、Br-、I-、CN-、PO4 3-、Ac-、SO4 2-、CO3 2-、HSO4 -、SCN-、HS-、SO3 2-、HCO3 -、NO3 -). As can be seen from FIGS. 12 and 13, ions other than hypochlorite ions have almost no fluorescence response to the fluorescent probe of the present invention, while hypochlorite ions have a significant fluorescence response to the fluorescent probe of the present invention, which indicates that the fluorescent probe of the present invention has a high selectivity for detection of mercury ions and can be used for detection of hypochlorite ions.
B, taking out 30 mu L of the fluorescent probe solution prepared in the step a, adding the fluorescent probe solution into a test tube, and adding the fluorescent probe solution into the test tube at a concentration of 1.00 multiplied by 10-3mol·L -115 μ L of sodium hypochlorite standard solution, then diluted to 3mL with PBS, the color of the solution before and after reaction with hypochlorite ions changed under fluorescent light, as shown in fig. 14; it can be seen that, under the daylight lamp, the hypochlorite ions can make the PBS solution of the fluorescent probe undergo a significant color change, and the solution color changes from orange (fig. 14 (a)) to yellow-green (fig. 14 (b)); the color change of the solution before and after the reaction with hypochlorite ions under the ultraviolet lamp is shown in fig. 15, and the hypochlorite ions induce the fluorescent probe to emit yellow green fluorescence (fig. 15 (b)) visible under the ultraviolet lamp, which indicates that the fluorescent probe is a fluorescent probe with a color-generating sensing function, can be used for visual qualitative detection of hypochlorite ions, and greatly facilitates the rapid qualitative detection of hypochlorite ions.
Claims (8)
1. A fluorescent probe for detecting mercury ions and hypochlorite ions is characterized in that the molecular formula of the fluorescent probe is C19H19NO3S2Structural formula is asThe following:
2. the method for preparing a fluorescent probe for detecting mercury ions and hypochlorite ions as claimed in claim 1, wherein the synthetic route is as follows:
the method specifically comprises the following steps:
(1) adding 4-bromo-1, 8-naphthalic anhydride and n-butylamine into ethanol, refluxing under the protection of nitrogen until the reaction solution is clear, concentrating, cooling to separate out crystals, and recrystallizing to obtain a compound 1;
(2) dissolving the compound 1 prepared in the step (1) in methanol, adding sodium methoxide and copper sulfate, performing reflux reaction for 11-15 h, concentrating, cooling, precipitating crystals, and washing with deionized water to obtain a compound 2;
(3) adding the compound 2 prepared in the step (2) into HI, performing reflux reaction for 12-24 h, cooling, filtering and washing to obtain a compound 3;
(4) adding the compound 3 prepared in the step (3), urotropine and paraformaldehyde into acetic acid, carrying out reflux reaction for 1-3 h, cooling to 80-90 ℃, adding excessive concentrated hydrochloric acid, continuing to react for 35-40 min, cooling, filtering to obtain filtrate, extracting with dichloromethane to obtain a crude product, and separating the crude product by using column chromatography to obtain a compound 4;
(5) adding the compound 4 prepared in the step (4), anhydrous magnesium sulfate and p-toluenesulfonic acid into tetrahydrofuran, and stirring under the protection of nitrogen to obtain a mixed solution; dissolving 1, 4-ethanedithiol in tetrahydrofuran, adding the tetrahydrofuran into the mixed solution, stirring at normal temperature for 6-8 h, removing the solvent after the reaction is finished, and performing column chromatography separation to obtain the fluorescent probe.
3. The method for preparing the fluorescent probe for detecting mercury ions and hypochlorite ions according to claim 2, wherein the molar ratio of 4-bromo-1, 8-naphthalic anhydride to n-butylamine in step (1) is 1 (5-7).
4. The method for preparing the fluorescent probe for detecting mercury ions and hypochlorite ions according to claim 2, wherein the molar ratio of the compound 1 to sodium methoxide in the step (2) is 1 (8-12).
5. The method for preparing a fluorescent probe for detecting mercury ions and hypochlorite ions according to claim 2, wherein the amount of the compound 2 in the step (3) is 1.5-2.0 g, and the amount of HI is 20-40 mL.
6. The method for preparing the fluorescent probe for detecting mercury ions and hypochlorite ions according to claim 2, wherein the molar ratio of the compound 3 to the urotropine in the step (4) is 1 (1-1.5); the mass ratio of the urotropine to the paraformaldehyde is 1 (1-1.5).
7. The method for preparing a fluorescent probe for detecting mercury ions and hypochlorite ions according to claim 2, wherein the molar ratio of the compound 4, anhydrous magnesium sulfate, p-toluenesulfonic acid and 1, 4-ethanedithiol in the step (5) is 1 (1-3) to (1-1.2).
8. The use of the fluorescent probe of claim 1 in the fluorescent detection of mercury ions and hypochlorite ions for fluorescence detection, visual qualitative detection and cellular imaging detection of mercury ion and hypochlorite ion content.
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