Pyridoxal disulfide Hg < 2 + > fluorescent probe and preparation method thereof
By designing a pyridoxal disulfide Hg2+ fluorescent probe, the existing Hg2+ detection methods are solved, and the detection effect of high selectivity, sensitivity and anti-interference ability is achieved. Trace analysis of Hg2+ in water and environment.
Patent Information
- Application Number
- CN202411423283.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-12
- Publication Date
- 2025-02-18
AI Technical Summary
The existing Hg2+ detection methods have disadvantages such as weak selectivity, susceptibility to environmental factors and difficulty in storage, making it difficult to achieve convenient and sensitive, highly selective and colorimetric recognition detection effects.
Design and develop a fluorescent probe of pyridoxal disulfide Hg2+, through the substitution reaction of α-bromoacetophenone and ammonium pyrrolidine dithioformate, the acylation reaction of the intermediate with hydrazine hydrate, and finally with pyridoxal The amine aldehyde condensation of hydrochloride salts can produce fluorescent probes with high selectivity and sensitivity.
It realizes specific identification of Hg2+, has high sensitivity and anti-interference ability, has a wide range of application, can show good detection effect within a wide pH range, has short response time, no secondary pollution, and is suitable for water and environment Trace analysis and detection of Hg2+.
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Figure CN119462495A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to a pyridoxal disulfide Hg 2+ The invention discloses a fluorescent probe and a preparation method thereof, specifically belonging to the field of organic functional materials. Background Art
[0002] Mercury is a transition heavy metal with high density, silvery white color, and liquid at room temperature. It is ubiquitous in nature. Some animals and plants contain trace amounts of mercury, which can be slowly metabolized through excretion, hair, etc. Because mercury exists in liquid form at room temperature and pressure, has stable chemical properties, and is insoluble in acids and alkalis, mercury and its compounds are widely used in the electronics manufacturing industry, chemical industry, metallurgical casting, medical treatment, and military. However, mercury is a typical global pollutant. It not only exists stably in the atmosphere, but can also enter the water and soil environment through atmospheric dry and wet deposition, thereby causing systemic damage to the ecosystem. Therefore, mercury pollution can easily threaten environmental safety and human health. Because mercury evaporates slowly and is not easily degraded, its impact on human health is irreversible. When mercury is slowly absorbed by the human body, it will be distributed to tissues throughout the body through the blood, damaging the central and peripheral nervous systems and organs such as the kidneys, and causing acute and chronic mercury poisoning. With the development of global industrialization, research on Hg in the environment and water bodies has been conducted. 2+ It is particularly important to develop efficient and convenient identification methods to obtain specificity, sensitivity and achieve trace detection.
[0003] Hg 2+ The detection methods mainly include electrochemical analysis, capillary electrophoresis, etc. These detection methods usually require long testing time, expensive and complex instruments, high testing requirements and low sensitivity. In recent years, colorimetric and fluorescence detection and analysis technologies have become the most popular methods for detecting Hg due to their advantages such as rapidity, sensitivity, ease of operation and real-time monitoring. 2+ Currently, many types of Hg 2+ Probes, such as nanoparticles and polymers, are mostly weak in selectivity, easily interfered by environmental factors, and difficult to store, which limits their application in environmental and biological analysis. Pyridoxal is one of the components of vitamin B6. Pyridoxal compounds and their derivatives have good biocompatibility. The synthesis of hydrazide Schiff base compounds based on pyridoxal can provide suitable sites to identify target ions, which are not interfered by environmental factors and can be identified by naked eyes and used for on-site detection. Therefore, the design and development of Hg 2+ The realization of convenient, sensitive, highly selective and colorimetric fluorescent probes is of great significance and practical application value. Summary of the invention
[0004] In view of the above situation, the purpose of the present invention is to provide a pyridoxal disulfide Hg 2+Fluorescent probe and preparation method. The fluorescent probe of the present invention can be used to detect Hg in water and natural environment. 2+ It has the characteristics of good selectivity, high sensitivity, simple operation, strong anti-interference ability and wide pH application range.
[0005] The present invention is a pyridoxal disulfide Hg 2+ The chemical structure of the fluorescent probe is: ; The present invention is a pyridoxal disulfide Hg 2+ The preparation method of the fluorescent probe is to firstly carry out a substitution reaction between α-bromoacetophenone and ammonium pyrrolidine dithiocarboxylate, and then carry out an acylation reaction between the intermediate 1 and hydrazine hydrate, and then carry out an amine-aldehyde condensation reaction between the intermediate 2 and pyridoxal hydrochloride to obtain pyridoxal disulfide Hg 2+ The fluorescent probe specifically comprises the following steps: Step 1: Under argon protection, α-bromoacetophenone and ammonium pyrrolidine dithiocarbamate are reacted in an acetone solution at room temperature for 2-4 h. The product is filtered off under reduced pressure to remove acetone, and then washed with anhydrous ethanol and dried to obtain intermediate 1; Step 2: The intermediate 1 and hydrazine hydrate are heated to reflux in a mixed solvent of tetrahydrofuran and anhydrous ethanol for 2-3 h. The product is cooled to room temperature, washed with water and separated by column chromatography. The eluate is concentrated by rotary evaporation to obtain the intermediate 2. Step 3: Intermediate 2, pyridoxal hydrochloride and sodium acetate were heated to reflux in anhydrous ethanol for 4-6 h, and a light yellow confetti-like precipitate was precipitated, which was filtered and washed with ethanol to obtain the product pyridoxal disulfide Hg 2+ Fluorescent probe (BDS).
[0006] In the step 1, the feed ratio of α-bromoacetophenone and ammonium pyrrolidinedithiocarbamate is 1 mmol:1 mmol.
[0007] In the step 2, the molar ratio of the intermediate 1 to hydrazine hydrate is 1:10, the volume ratio of the mixed solvent of tetrahydrofuran and anhydrous ethanol is 1:1, and the eluent used for column chromatography separation is ethyl acetate and petroleum ether in a volume ratio of 3:7.
[0008] In the step 3, the molar ratio of intermediate 2, pyridoxal hydrochloride and sodium acetate is 1:1:4.
[0009] Reaction equation of the present invention: ; Where: 1 represents intermediate 1; 2 represents intermediate 2; BDS represents pyridoxal disulfide Hg 2+ Fluorescent probe compounds.
[0010] The present invention pyridoxal disulfide Hg2+ How to use fluorescent probes: Pyridoxal disulfide Hg 2+ The fluorescent probe is used by 2+ The aqueous solution of Hg was added dropwise to the DMSO (dimethyl sulfoxide) / H2O (v / v, 8:2) mixed solution of the probe molecule BDS. During the process, the solution was observed to change from orange-red fluorescence to yellow-white fluorescence. 2+ No fluorescence change occurs in solutions of other metal cations except
[0011] The specific operation is as follows: add appropriate amount of DMSO / H2O (v / v, 8:2) solvent to 19 5 mL centrifuge tubes, and then drop Li + , Na + ,Mg 2+ , Al 3+ , K + , Ca 2+ ,Cr 3+ , Mn 2+ , Fe 2+ , Fe 3+ ,Co 2+ , Ni 2+ , Zn 2+ , Ag + , Cd 2+ , Ba 2+ , Pb 2+ , Ce 3+ and Hg 2+ Finally, the probe molecule BDS dilute solution was added dropwise, and Hg 2+ The fluorescence of the aqueous solution in the centrifuge tube changed from orange-red to yellow-white, while other metal cation solutions showed no fluorescence change.
[0012] Beneficial effects of the present invention: 1. The present invention adopts a three-step simple reaction to combine the disulfide functional group-containing component with the pyridoxal moiety. The raw materials are relatively cheap, the reaction conditions are mild, the preparation process is simple, and the yield is high.
[0013] 2. Pyridoxal disulfide Hg of the present invention 2+ The molecular structure of the fluorescent probe is synthesized for the first time, with good solubility, strong anti-interference ability, specific recognition, high sensitivity, and can effectively distinguish metal Hg 2+ and other common metal ions.
[0014] 3. Pyridoxal disulfide Hg of the present invention 2+The fluorescent probe has a wide range of applications and can show good detection effects in a wide pH range. The luminescent wavelengths are 494 nm and 518 nm, respectively, which are yellow-white fluorescence visible to the naked eye. It has a short response time and will not produce secondary pollution during use, which is beneficial to the detection of Hg in water and the environment. 2+ Trace analysis and detection has important application prospects and promotion value. BRIEF DESCRIPTION OF THE DRAWINGS
[0015] Figure 1 : Pyridoxal disulfide Hg of the present invention 2+ Chemical structure of the fluorescent probe; Figure 2 : Pyridoxal disulfide Hg of the present invention 2+ H NMR spectrum of fluorescent probe ( 1 H-NMR) spectrum; Figure 3 : Pyridoxal disulfide Hg of the present invention 2+ Fluorescent probe for Hg in DMSO / H2O system 2+ Graph of fluorescence changes in response; Figure 4-1 : Pyridoxal disulfide Hg of the present invention 2+ Fluorescent probes for different concentrations of Hg in DMSO / H2O system 2+ Fluorescence emission spectrum of Figure 4-2 : Pyridoxal disulfide Hg of the present invention 2+ The fluorescence enhancement degree of the fluorescent probe in the DMSO / H2O system is related to Hg 2+ The relationship between concentrations; Figure 5 : Pyridoxal disulfide Hg of the present invention 2+ Fluorescence probe probe molecule BDS and Hg in DMSO / H2O system 2+ Combined Job graph; Figure 6 : Pyridoxal disulfide Hg of the present invention 2+ Detection of the anti-interference ability of fluorescent probes in the presence of different metal ions; Figure 7 : Pyridoxal disulfide Hg of the present invention 2+ Fluorescence intensity changes of fluorescent probe under different pH conditions.
[0016] Figure 8-1 : Pyridoxal disulfide Hg of the present invention 2+ Fluorescence emission spectra of fluorescent probes at different response times; Figure 8-2 : Pyridoxal disulfide Hg of the present invention 2+ The relationship between the fluorescence intensity and response time of the fluorescent probe. DETAILED DESCRIPTION
[0017] The present invention is described in detail below in conjunction with the accompanying drawings, but it should be noted that implementation examples of the present invention include but are not limited to the following implementation methods.
[0018] Example 1 Synthesis of Pyridoxal Disulfide Hg2+ Probe Synthesis of intermediate 1: α-Bromoacetophenone (0.5892 g, 2.98 mmol) and ammonium pyrrolidine dithiocarboxylate (0.4895 g, 2.98 mmol) were dissolved in 6 mL of acetone solvent, protected by argon atmosphere, and stirred at room temperature until the reaction was completed (the reaction progress was monitored by TLC thin-layer chromatography). After the reaction was completed, the product was filtered under reduced pressure, rinsed three times with a small amount of ethanol, and the solid was dried and weighed to obtain 0.6772 g of light yellow solid powder intermediate 1, with a yield of 85.9%.
[0019] Synthesis of intermediate 2: The intermediate 1 (0.6772 g, 2.55 mmol) was added to a mixed solvent of tetrahydrofuran / anhydrous ethanol (v / v, 5 mL: 5 mL), and then 1.5 mL of hydrazine hydrate (mass concentration of 40% to 50%) was added. The oil bath temperature was set to 85°C under argon atmosphere and heated to reflux for 3 h until the reaction was completed (TLC thin layer chromatography was used to monitor the reaction progress). The reactants were cooled to room temperature, and the crude product was separated by column chromatography (eluent: ethyl acetate: petroleum ether = 3:7, volume ratio), and a light brown oily intermediate 2 was obtained by rotary evaporation under reduced pressure on a rotary evaporator with a yield of 71.2%.
[0020] Synthesis of probe molecule BDS: Pyridoxal hydrochloride (0.1093 g, 0.65 mmol), organic intermediate 2 (0.1814 g, 0.65 mmol) and sodium acetate (0.2133 g, 2.6 mmol) were sequentially added into 10 mL of anhydrous ethanol solvent, the oil bath temperature was set at 78 °C, and the mixture was heated under reflux for 5 h until the reaction was completed (TLC monitoring). The reaction solution was cooled to room temperature, and a light yellow confetti-like precipitate was precipitated. The mixture was filtered under reduced pressure and rinsed three times with a small amount of ethanol to obtain a light yellow pyridoxal disulfide Hg 2+ Fluorescent probe BDS (0.1897 g, yield 68.1%).
[0021] Pyridoxal disulfide Hg 2+ The H NMR spectrum of the fluorescent probe is Figure 2 shown. 1H NMR (500 MHz, DMSO-d6) δ = 11.65 (s, 1H), 9.14 (s, 1H), 8.03 (s, 1H), 8.01 - 7.96 (m, 2H), 7.59- 7.48 (m, 3H), 5.48 (t, J = 5.4 Hz, 1H), 4.87 (s, 2H), 4.70 (d, J = 4.7 Hz,2H), 3.79 (t, J = 6.9 Hz, 2H), 3.54 (t, J = 6.8 Hz, 2H), 2.44 (s, 3H), 1.96(q, J = 6.7 Hz, 2H), 1.90 (q, J = 6.8 Hz, 2H). The spectrum shows that the pyridoxal disulfide Hg prepared by the present invention 2+ The fluorescent probe has the same structure as the target molecule.
[0022] Example 2 Pyridoxal disulfide Hg 2+ Fluorescent probe for Hg 2+ Selective detection of fluorescence responses.
[0023] DMSO / H2O (v / v, 8:2) system was used as the experimental test environment.
[0024] Pyridoxal disulfide Hg 2+ The fluorescent probe was dissolved in DMSO solvent and diluted to 50 mL in a volumetric flask to prepare a standard solution with a concentration of 1 mmol / L as the ion detection solution.
[0025] Take 20 5 mL centrifuge tubes and add 2.25 mL of DMSO solvent to each centrifuge tube. The first centrifuge tube is used as a blank control group. Add 0.6 mL of 1 mmol / L Li-containing solution to the remaining 19 centrifuge tubes. + , Na + ,Mg 2+ , Al 3 + , K + , Ca 2+ ,Cr 3+ , Mn 2+ , Fe 2+ , Fe 3+ ,Co 2+ , Ni 2+ , Zn2+ , Ag + , Cd 2+ , Ba 2+ , Pb 2+ , Ce 3+ and Hg 2+ Aqueous solution, and finally add 0.15 mL of ion detection solution with a concentration of 1 mmol / L. The above sample solution was allowed to stand for 30 min, and then transferred to a standard quartz cuvette to measure its fluorescence spectrum change curve. The excitation wavelength was set to 421 nm, and the maximum emission wavelengths were set to 494 nm and 518 nm.
[0026] Pyridoxal disulfide Hg 2+ Fluorescent probe for Hg 2+ The fluorescence response effect is as follows Figure 3 The results showed that the probe molecule BDS and Hg 2+ The combination shows obvious fluorescence enhancement at 494 nm and 518 nm. The research results show that the pyridoxal disulfide Hg 2+ Fluorescent probe for Hg 2+ It has a specific fluorescence enhancement effect with high sensitivity.
[0027] Example 3 Pyridoxal disulfide Hg 2+ Fluorescent probe for Hg 2+ Quantitative fluorescence detection.
[0028] DMSO / H2O (v / v, 8:2) system was used as the experimental test environment.
[0029] Take 10 5 mL centrifuge tubes, add 2.3 mL of DMSO solvent, 0.05-0.5 mL of deionized water, and then add 1 eq - 6 eq of 1 mmol / L Hg 2+ (0.1-0.55 mL) aqueous solution, and finally add 0.1 mL of 1 mmol / L sample test solution to each centrifuge tube. Let the above samples stand for 30 minutes, then transfer them to a standard quartz cuvette and measure the changes in their fluorescence spectra.
[0030] Pyridoxal disulfide Hg 2+ Fluorescent probes for different concentrations of Hg 2+ The fluorescence emission spectrum of Figure 4-1 The fluorescence intensity value at 494 nm of the fluorescence emission spectrum is compared with the corresponding Hg 2+ The standard curve was obtained by fitting the concentration equivalent ratio (such as Figure 4-2 As shown), the results show that the linear relationship is good, indicating that the pyridoxal disulfide Hg2+ Fluorescent probe can quantitatively detect Hg in DMSO / H2O system 2+ concentration.
[0031] Example 4 Probe molecule BDS and Hg in solution system 2+ complexation ratio.
[0032] DMSO / H2O (v / v, 8:2) system was used as the experimental test environment.
[0033] Take 9 5 mL centrifuge tubes and control the probe BDS and Hg 2+ The total concentration of Hg was 200 μM. The probe molecule BDS solution / Hg 2+ The molar concentration ratio of the aqueous solution was 1:9, 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, 9:1, and then DMSO solvent (1.86-2.34 mL) and deionized water (0.06-0.54 mL) were added respectively, and the total volume was kept constant at 3 mL. The above samples were allowed to stand for 30 minutes, and then transferred to a standard quartz cuvette to measure their fluorescence value at 494 nm.
[0034] Pyridoxal disulfide Hg 2+ Fluorescence probe probe molecule BDS and Hg in DMSO / H2O system 2+ The combined Job curve is as follows Figure 5 The results show that the fluorescence value is [BDS] / {[BDS]+[Hg 2+ ]} appears an inflection point when it is 0.5, indicating that the pyridoxal disulfide Hg 2+ Fluorescence probe probe molecule BDS and Hg in DMSO / H2O system 2+ The complexation ratio is 1:1.
[0035] Example 5 Pyridoxal disulfide Hg 2+ Detection of the anti-interference ability of fluorescent probes.
[0036] DMSO / H2O (v / v, 8:2) system was used as the experimental test environment.
[0037] Take 40 5 mL centrifuge tubes and divide them into two groups, with 20 sample tubes in each group. In the first group, 2.34 mL of DMSO solvent and 0.3 mL of deionized water were added to each sample tube. The first sample tube was used as a blank control group. The remaining 19 centrifuge tubes were added with 0.3 mL of 1 mmol / L Li-containing + , Na + ,Mg 2+, Al 3+ , K + , Ca 2+ ,Cr 3+ , Mn 2+ , Fe 2+ , Fe 3+ ,Co 2+ , Ni 2+ , Zn 2+ , Ag + , Cd 2+ , Ba 2+ , Pb 2+ , Ce 3+ and Hg 2+ In the second group, 2.34 mL of DMSO solvent was added to each sample tube, and 0.3 mL of 1 mmol / LHg was added to each sample tube. 2+ The first sample tube was used as a blank control group, and 0.3 mL of 1 mmol / L Li-containing + , Na + ,Mg 2+ , Al 3 + , K + , Ca 2+ ,Cr 3+ , Mn 2+ , Fe 2+ , Fe 3+ ,Co 2+ , Ni 2+ , Zn 2+ , Ag + , Cd 2+ , Ba 2+ , Pb 2+ , Ce 3+ and Hg 2+ Aqueous solution, and finally add 0.06 mL of 1 mmol / L ion detection solution. After the above sample solution is allowed to stand for 30 minutes, it is transferred to a standard quartz cuvette and its fluorescence intensity value at 494 nm is measured.
[0038] Pyridoxal disulfide Hg 2+ The anti-interference ability of fluorescent probes is Figure 6 The results show that without adding Hg 2+ The fluorescence value of the solution is very weak. When Hg 2+ After that, the fluorescence was significantly enhanced. This result shows that the pyridoxal disulfide Hg 2+ The fluorescent probe has strong anti-interference ability in the presence of different metal ions.
[0039] Example 6 pH for pyridoxal disulfide Hg 2+ The influence of fluorescent probe recognition performance.
[0040] DMSO / H2O (v / v, 8:2) system was used as the experimental test environment.
[0041] Eleven groups of aqueous solutions with different pH values in the pH range of 2-12 were prepared as pH adjusters.
[0042] Take 22 5 mL centrifuge tubes and divide them into two groups, each with 11 sample tubes. In the first group, add 2.25 mL of DMSO solvent, 0.45 mL of pH regulator with a pH of 2-12, and 0.15 mL of Hg with a concentration of 1 mmol / L to each sample tube. 2+ Aqueous solution, 0.15 mL of 1 mmol / L ion detection solution. In the second group, each sample tube was added with 2.25 mL of DMSO solvent, 0.6 mL of pH adjuster with a pH of 2-12, and 0.15 mL of 1 mmol / L ion detection solution. The sample solution was allowed to stand for 30 min, transferred to a standard quartz cuvette, and its fluorescence value at 494 nm was measured.
[0043] Pyridoxal disulfide Hg 2+ The fluorescence intensity of the fluorescent probe changes under different pH conditions. Figure 7 The results showed that the BDS probe itself exhibited weak fluorescence changes in the pH range of 2-12. 2+ After binding, a strong fluorescence change is shown in the pH range of 4-10. The results show that the pyridoxal disulfide Hg 2+ Fluorescent probes can be used for Hg in a wide range of pH conditions 2+ Detection.
[0044] Example 7 Pyridoxal disulfide Hg 2+ Response time determination of fluorescent probes.
[0045] DMSO / H2O (v / v, 8:2) system was used as the experimental test environment.
[0046] Take a 1 mL centrifuge tube and add 2.34 mL of DMSO solvent, 0.3 mL of deionized water, and 0.3 mL of 1 mmol / L Hg 2+Aqueous solution. Transfer the solution in the above sample tube to a standard quartz cuvette, add 0.06 mL of 1 mmol / L ion detection solution, and immediately place the cuvette into a fluorescence spectrometer for detection. Measure its fluorescence value every 2 minutes to obtain a time-fluorescence value change spectrum.
[0047] Pyridoxal disulfide Hg 2+ The fluorescence emission spectra of the fluorescent probe at different times are as follows Figure 8-1 The fluorescence intensity value at 494 nm of the fluorescence emission spectrum is fitted with the response time to obtain a relationship curve (as shown in Figure 8-2 As shown), the results show that the pyridoxal disulfide Hg prepared by the present invention 2+ The fluorescence intensity of the fluorescent probe increased with time and reached a peak value at 40 min.
Claims
1. A pyridoxal disulfide Hg 2+ A fluorescent probe, characterized in that: The pyridoxal disulfide Hg 2+ The chemical structure of the fluorescent probe is: 。 2. A pyridoxal disulfide Hg 2+ A method for preparing a fluorescent probe, characterized in that: The preparation method is to firstly carry out a substitution reaction between α-bromoacetophenone and ammonium pyrrolidine dithiocarboxylate, then carry out an acylation reaction between the intermediate 1 and hydrazine hydrate, and then carry out an amine-aldehyde condensation reaction between the intermediate 2 and pyridoxal hydrochloride to obtain pyridoxal disulfide Hg 2+ The fluorescent probe specifically comprises the following steps: Step 1: Under argon protection, α-bromoacetophenone and ammonium pyrrolidine dithiocarbamate are reacted in an acetone solution at room temperature for 2-4 h. The product is filtered off under reduced pressure to remove acetone, and then washed with anhydrous ethanol and dried to obtain intermediate 1; Step 2: The intermediate 1 and hydrazine hydrate are heated to reflux in a mixed solvent of tetrahydrofuran and anhydrous ethanol for 2-3 h. The product is cooled to room temperature, washed with water and separated by column chromatography. The eluate is concentrated by rotary evaporation to obtain the intermediate 2. Step 3: Intermediate 2, pyridoxal hydrochloride and sodium acetate were heated to reflux in anhydrous ethanol for 4-6 h, and a light yellow confetti-like precipitate was precipitated, which was filtered and washed with ethanol to obtain the product pyridoxal disulfide Hg 2+ Fluorescent probes.
3. A pyridoxal disulfide Hg according to claim 2 2+ A method for preparing a fluorescent probe, characterized in that: In the step 1, the feed ratio of α-bromoacetophenone and ammonium pyrrolidinedithiocarbamate is 1 mmol:1 mmol.
4. A pyridoxal disulfide Hg according to claim 2 2+ The method for preparing a fluorescent probe is characterized by: In the step 2, the molar ratio of the intermediate 1 to hydrazine hydrate is 1:10, the volume ratio of the mixed solvent of tetrahydrofuran and anhydrous ethanol is 1:1, and the eluent used for column chromatography separation is ethyl acetate and petroleum ether in a volume ratio of 3:
7.
5. A pyridoxal disulfide Hg according to claim 2 2+ The method for preparing a fluorescent probe is characterized by: In the step 3, the molar ratio of intermediate 2, pyridoxal hydrochloride and sodium acetate is 1:1:4.