Synthesis method and application of fluorescent probe for detecting hydrogen sulfide

Abstract

The invention relates to the technical field of chemical analysis and detection, and discloses a synthesis method and application of a fluorescent probe for detecting hydrogen sulfide. The fluorescence emission unit is selected from at least one or a combination of more of the following structures: carboxyl fluorescein, rhodamine B or other fluorescent dyes with high quantum yield and good water solubility; the hydrogen sulfide recognition group is selected from at least one of the following structures: a mercaptan compound with reactivity, an amino acid derivative or other compounds capable of forming a specific reaction with hydrogen sulfide. The fluorescent dye with high quantum yield and good water solubility, such as carboxyl fluorescein and rhodamine B, is adopted to ensure that the fluorescent dye emits bright and stable fluorescence in a wavelength range of 470nm to 650nm. Due to the characteristic of high quantum yield, the probe has excellent performance in optical imaging and fluorescence detection, and the visibility of the probe in low-concentration hydrogen sulfide detection is enhanced.

Description

Technical Field

[0001] The invention relates to the technical field of chemical analysis and detection, in particular to a synthesis method of a fluorescent probe for detecting hydrogen sulfide and application thereof. Background Art

[0002] Hydrogen sulfide (H2S) is a toxic and corrosive gas. Its presence in industrial emissions, natural gas, and sewage treatment may pose a serious threat to the environment and human health. Therefore, the development of efficient and reliable methods for hydrogen sulfide detection is of great practical significance. At present, the existing hydrogen sulfide detection technologies mainly include chemical sensors, gas chromatography, mass spectrometry, and spectroscopy. Among them, chemical sensors have attracted widespread attention due to their portability and real-time detection capabilities, but their sensitivity and selectivity are often limited. Although gas chromatography and mass spectrometry have high sensitivity and accuracy, their application in field detection is limited due to the expensive equipment and complex operation.

[0003] As a new type of sensor, fluorescent probes are favored by researchers because of their high sensitivity, rapid response and visualization. Traditional fluorescent probes usually use simple fluorescent dyes, but they often face problems such as insufficient fluorescence intensity and background noise interference when detecting low concentrations of hydrogen sulfide. In addition, the synthesis method of existing fluorescent probes requires multiple reaction steps, including esterification, condensation, redox reactions, etc. Each step requires specific reaction conditions, such as temperature, time, pressure, etc., which increases the complexity of the synthesis process and the synthesis efficiency is relatively low. At the same time, the lack of good solubility in aqueous phase affects its application in biological sample detection.

[0004] Therefore, those skilled in the art have proposed a method for synthesizing a fluorescent probe for detecting hydrogen sulfide and its application to solve the above problems. Summary of the invention

[0005] In view of the deficiencies of the prior art, the present invention provides a method for synthesizing a fluorescent probe for detecting hydrogen sulfide and its application, which solves the problems raised in the above-mentioned background technology.

[0006] To achieve the above objectives, the present invention is implemented by the following technical scheme: a fluorescent probe for detecting hydrogen sulfide, the chemical structure of the fluorescent probe is composed of the following components:

[0007] A fluorescence emission unit, wherein the fluorescence emission unit is selected from at least one or a combination of the following structures: carboxyfluorescein, rhodamine B or other fluorescent dyes with high quantum yield and good water solubility;

[0008] A hydrogen sulfide recognition group, wherein the recognition group is at least one selected from the following structures: a reactive thiol compound, an amino acid derivative or other compound capable of forming a specific reaction with hydrogen sulfide;

[0009] A connecting group connects the fluorescent emission unit and the hydrogen sulfide recognition group through a covalent bond, and the connecting group is selected from at least one of the following compounds: polyvinyl alcohol, polyethylene imine and polyacrylic acid.

[0010] Preferably, the molecular structure of the fluorescence emission unit emits fluorescence within a wavelength range of 470 nm to 650 nm, and the fluorescence quantum yield of the fluorescence emission unit is ≥0.5.

[0011] Preferably, the hydrogen sulfide recognition group reacts with hydrogen sulfide through a chemical reaction or a coordination reaction, causing a significant change in the fluorescence intensity of the probe.

[0012] Preferably, the probe can detect hydrogen sulfide in a concentration range of 0.01 μM to 10 mM when the environmental pH value is 6.0 to 8.0, and there is a good linear relationship between the fluorescence intensity and the hydrogen sulfide concentration.

[0013] A method for synthesizing a fluorescent probe for detecting hydrogen sulfide, the method comprising the following steps:

[0014] S1, under an inert gas atmosphere, mixing the fluorescent emission unit and the hydrogen sulfide recognition group in an organic solvent, wherein the solvent is deionized water, dimethyl sulfoxide or ethanol;

[0015] S2, heating the mixture to 50°C-80°C for a reaction time of 2h-8h to form a preliminary product;

[0016] S3, after the reaction is completed, adding a linking group and continuing the heating reaction for 4h-24h to obtain the final fluorescent probe;

[0017] S4, preferably purifying the fluorescent probe by dialysis, liquid chromatography or other separation techniques.

[0018] Preferably, in step S3, the connecting group is polyvinyl alcohol, which is connected to the fluorescent emission unit and the recognition group through an esterification reaction, and the reaction temperature is carried out at room temperature.

[0019] The invention discloses an application of a fluorescent probe for detecting hydrogen sulfide. The probe can be used for detecting hydrogen sulfide in cell biology research, including monitoring of hydrogen sulfide concentration inside and outside cells and its role as a biomarker in diseases.

[0020] Preferably, the fluorescent probe can monitor the dynamic changes of hydrogen sulfide concentration in real time in a cell model, and its sensitivity reaches 0.1 μM, has good selectivity, and is not interfered by other biological molecules.

[0021] Preferably, the probe can be used for environmental monitoring, especially for real-time monitoring of hydrogen sulfide concentration in water quality monitoring, air quality assessment and soil pollution monitoring.

[0022] The invention provides a synthesis method of a fluorescent probe for detecting hydrogen sulfide and application thereof.

[0023] It has the following beneficial effects:

[0024] 1. The present invention uses fluorescent dyes with high quantum yield and good water solubility such as carboxyfluorescein and rhodamine B to ensure that it emits bright and stable fluorescence in the wavelength range of 470nm to 650nm. This high quantum yield property enables the probe to perform well in optical imaging and fluorescence detection, enhances the visibility of the probe in low-concentration hydrogen sulfide detection, and thus improves the accuracy and reliability of the detection.

[0025] 2. The synthesis method of the fluorescent probe of the present invention is simple and efficient, involving mixing, heating reaction and subsequent purification steps under an inert gas atmosphere. By mixing the fluorescent emission unit with the hydrogen sulfide recognition group and then heating the reaction, and then introducing the linking group, the efficient synthesis of the fluorescent probe is achieved. The use of polyvinyl alcohol not only improves the stability of the probe, but also enhances its solubility in the aqueous phase, making it suitable for the detection of biological samples. This efficient synthesis method can reduce production costs and promote large-scale applications.

[0026] 3. The fluorescent probe prepared by the present invention can efficiently detect hydrogen sulfide in a concentration range of 0.01 μM to 10 mM when the environmental pH value is 6.0 to 8.0. There is a good linear relationship between its fluorescence intensity and hydrogen sulfide concentration, which makes it possible to use the probe in different biological samples and environmental samples. In particular, in cell biology research, the probe can monitor the dynamic changes of hydrogen sulfide concentration in real time with a sensitivity of 0.1 μM, providing a reliable tool for biomarker detection in disease research. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 It is a process flow chart of the present invention. DETAILED DESCRIPTION

[0028] The following will be combined with the drawings in the specification of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] Embodiment 1:

[0030] Please see attached Figure 1 The embodiment of the present invention provides a fluorescent probe for detecting hydrogen sulfide. The chemical structure of the fluorescent probe is composed of the following components:

[0031] A fluorescence emission unit, wherein the fluorescence emission unit is selected from at least one or a combination of the following structures: carboxyfluorescein, rhodamine B or other fluorescent dyes with high quantum yield and good water solubility;

[0032] Specifically, the molecular structure of the fluorescence emission unit can emit fluorescence within the wavelength range of 470nm, ensuring its effectiveness in the visible spectrum and facilitating matching with light sources and detectors. In addition, the requirement of fluorescence quantum yield ≥ 0.5 ensures the sensitivity of the probe in practical applications and can effectively respond to the presence of target molecules.

[0033] A hydrogen sulfide recognition group, wherein the recognition group is at least one selected from the following structures: a reactive thiol compound, an amino acid derivative or other compound capable of forming a specific reaction with hydrogen sulfide;

[0034] Specifically, the hydrogen sulfide recognition group is another key part of the probe, responsible for selectively recognizing and responding to hydrogen sulfide (H2S). The recognition groups used in this probe design include:

[0035] Reactive thiol compounds: These compounds usually react rapidly with hydrogen sulfide to form stable derivatives, which leads to changes in the fluorescence properties of the probe.

[0036] Amino acid derivatives: Specific amino acid derivatives can form specific reactions with hydrogen sulfide through their side chains. This specificity improves the selectivity and sensitivity of the probe.

[0037] Other compounds that can form specific reactions with hydrogen sulfide: such as certain functionalized metal complexes, which can form complexes with hydrogen sulfide through coordination reactions, further enhancing the response ability of the probe

[0038] The molecular structure of the fluorescence emission unit emits fluorescence within a wavelength range of 470 nm, and the fluorescence quantum yield of the fluorescence emission unit is ≥0.5.

[0039] The hydrogen sulfide recognition group reacts with hydrogen sulfide through chemical reaction or coordination reaction, causing the fluorescence intensity of the probe to change significantly. When the environmental pH value is 6.2, the probe can detect hydrogen sulfide in the concentration range of 0.01μM to 10mM, and there is a good linear relationship between the fluorescence intensity and the hydrogen sulfide concentration.

[0040] Specifically, the probe has excellent performance at an environmental pH of 6.2, and can accurately detect hydrogen sulfide in a concentration range of 0.01 μM to 10 mM. This range is designed to take into account the actual conditions of biological samples, making the probe have good application prospects both in vivo and in the external environment.

[0041] Based on the above characteristics, this fluorescent probe has broad application prospects in many fields:

[0042] Biological imaging: It can be used for in vivo and in vitro biological imaging to help researchers observe the distribution of hydrogen sulfide in organisms and its biological effects.

[0043] Environmental monitoring: Rapidly detect hydrogen sulfide concentration in water or gas to assess the degree of environmental pollution.

[0044] Medical diagnosis: used for early diagnosis of diseases, such as detection of relevant biomarkers of cardiovascular disease or neurodegenerative diseases.

[0045] A method for synthesizing a fluorescent probe for detecting hydrogen sulfide, the method comprising the following steps:

[0046] S1, under an inert gas atmosphere, mixing the fluorescent emission unit and the hydrogen sulfide recognition group in an organic solvent, the solvent being deionized water, dimethyl sulfoxide or ethanol;

[0047] S2, heating the mixture to 50°C for 2 h to form a preliminary product;

[0048] S3, after the reaction is completed, a linker group is added and the heating reaction is continued for 4 h to obtain the final fluorescent probe;

[0049] In step S3, the connecting group is polyvinyl alcohol, which is connected to the fluorescent emission unit and the recognition group through an esterification reaction, and the reaction temperature is carried out at room temperature.

[0050] S4, purify the fluorescent probe by dialysis, liquid chromatography or other separation techniques.

[0051] The connecting group connects the fluorescent emission unit and the hydrogen sulfide recognition group through a covalent bond, and the connecting group is selected from at least one of the following compounds: polyvinyl alcohol, polyethylene imine and polyacrylic acid.

[0052] An application of a fluorescent probe for detecting hydrogen sulfide, the probe can be used for hydrogen sulfide detection in cell biology research, including monitoring of hydrogen sulfide concentration inside and outside cells and its role as a biomarker in diseases. The fluorescent probe can monitor the dynamic changes of hydrogen sulfide concentration in real time in a cell model, and its sensitivity reaches 0.1μM, has good selectivity, and is not interfered by other biological molecules. The probe can be used for environmental monitoring, especially for real-time monitoring of hydrogen sulfide concentration in water quality testing, air quality assessment, and soil pollution monitoring.

[0053] Specifically, the synthesis method of the fluorescent probe is simple and efficient, involving mixing, heating reaction and subsequent purification steps under an inert gas atmosphere. The fluorescent probe is efficiently synthesized by mixing the fluorescent emission unit with the hydrogen sulfide recognition group and then heating the reaction, followed by the introduction of the linking group. The use of polyvinyl alcohol not only improves the stability of the probe, but also enhances its solubility in the aqueous phase, making it suitable for the detection of biological samples. This efficient synthesis method can reduce production costs and promote large-scale applications.

[0054] Embodiment 2 is different from Embodiment 1 in that:

[0055] Fluorescence emission unit: using fluorescein and acridine orange, with a high fluorescence quantum yield (≥0.6).

[0056] Hydrogen sulfide recognition group: N-ethyl amino acid is selected, which can effectively react with hydrogen sulfide.

[0057] Connecting group: Polyethyleneimine is used to connect the fluorescent emission unit and the recognition group through chemical bonds.

[0058] Detection range: 0.05μM to 5mM, with good linear relationship.

[0059] Embodiment 3 is different from Embodiment 1 in that:

[0060] Fluorescence emission unit: Select green fluorescent protein, which has a high fluorescence quantum yield (≥0.55).

[0061] Hydrogen sulfide recognition group: Use thiol compounds, which are highly reactive.

[0062] Connecting group: Connected with other components through polyacrylic acid by esterification reaction.

[0063] Detection range: 0.1μM to 1mM, showing good linear relationship.

[0064] Embodiment 4 is different from Embodiment 1 in that:

[0065] Fluorescence emission unit: using benzimidazole and rhodamine 6G, the fluorescence quantum yield is ≥ 0.7.

[0066] Hydrogen sulfide recognition group: Using amino acid derivatives with affinity can effectively recognize hydrogen sulfide.

[0067] Linking group: polyvinyl alcohol, using heating reaction and dialysis purification.

[0068] Detection range: 0.01μM to 20mM, maintaining good linear relationship.

[0069] The following table 1 is a detailed description and comparison of Examples 1 to 4:

[0070]

[0071]

[0072]

[0073] Conclusion: The fluorescence quantum yields of all examples are 0.5 and above, indicating that these probes have excellent performance in fluorescence emission efficiency and can effectively respond to changes in hydrogen sulfide.

[0074] Sensitivity: The detection range of each example is different, with the minimum detection limit ranging from 0.01 μM to 0.1 μM and the maximum detection limit reaching 20 mM. These differences in range reflect the diversity of different probes in sensitivity and application scenarios.

[0075] Quantitative analysis: All examples showed good linear relationships, indicating that there was a predictable relationship between the fluorescence intensity of the probe and the concentration of hydrogen sulfide, and were suitable for quantitative analysis.

[0076] In summary: The synthesis method of the fluorescent probe is simple and efficient, involving mixing, heating reaction and subsequent purification steps under an inert gas atmosphere. By mixing the fluorescent emission unit with the hydrogen sulfide recognition group and then heating the reaction, and then introducing the connecting group, the efficient synthesis of the fluorescent probe is achieved. At the same time, the fluorescent probe prepared by the present invention can efficiently detect hydrogen sulfide in a concentration range of 0.01μM to 10mM when the environmental pH value is 6.0 to 8.0. There is a good linear relationship between its fluorescence intensity and hydrogen sulfide concentration, which makes it possible to use the probe in different biological samples and environmental samples.

[0077] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A fluorescent probe for detecting hydrogen sulfide, characterized in that: The chemical structure of the fluorescent probe consists of the following components: A fluorescence emission unit, wherein the fluorescence emission unit is selected from at least one or a combination of the following structures: carboxyfluorescein, rhodamine B or other fluorescent dyes with high quantum yield and good water solubility; A hydrogen sulfide recognition group, wherein the recognition group is at least one selected from the following structures: a reactive thiol compound, an amino acid derivative or other compound capable of forming a specific reaction with hydrogen sulfide; A connecting group connects the fluorescent emission unit and the hydrogen sulfide recognition group through a covalent bond, and the connecting group is selected from at least one of the following compounds: polyvinyl alcohol, polyethylene imine and polyacrylic acid.

2. A fluorescent probe for detecting hydrogen sulfide according to claim 1, characterized in that: The molecular structure of the fluorescence emission unit emits fluorescence within a wavelength range of 470 nm to 650 nm, and the fluorescence quantum yield of the fluorescence emission unit is ≥0.

5.

3. A fluorescent probe for detecting hydrogen sulfide according to claim 1, characterized in that: The hydrogen sulfide recognition group reacts with hydrogen sulfide through a chemical reaction or a coordination reaction, causing a significant change in the fluorescence intensity of the probe.

4. A fluorescent probe for detecting hydrogen sulfide according to claim 1, characterized in that: The probe can detect hydrogen sulfide in a concentration range of 0.01 μM to 10 mM when the environmental pH value is 6.0 to 8.0, and there is a good linear relationship between the fluorescence intensity and the hydrogen sulfide concentration.

5. A method for synthesizing a fluorescent probe for detecting hydrogen sulfide, according to a fluorescent probe for detecting hydrogen sulfide according to any one of claims 1 to 4, characterized in that: The synthesis method The following steps are involved: S1, under an inert gas atmosphere, mixing the fluorescent emission unit and the hydrogen sulfide recognition group in an organic solvent, wherein the solvent is deionized water, dimethyl sulfoxide or ethanol; S2, heating the mixture to 50°C-80°C for a reaction time of 2h-8h to form a preliminary product; S3, after the reaction is completed, adding a linking group and continuing the heating reaction for 4h-24h to obtain the final fluorescent probe; S4, purifying the fluorescent probe by dialysis, liquid chromatography or other separation techniques.

6. The method for synthesizing a fluorescent probe for detecting hydrogen sulfide according to claim 5, characterized in that: In the step S3, the connecting group is polyvinyl alcohol, which is connected to the fluorescent emission unit and the recognition group through an esterification reaction, and the reaction temperature is carried out at room temperature.

7. Use of a fluorescent probe for detecting hydrogen sulfide, according to the fluorescent probe for detecting hydrogen sulfide according to any one of claims 1 to 4, characterized in that: The probe can be used for hydrogen sulfide detection in cell biology research, including monitoring of hydrogen sulfide concentration inside and outside cells and its role as a biomarker in diseases.

8. The use of a fluorescent probe for detecting hydrogen sulfide according to claim 7, characterized in that: The fluorescent probe can monitor the dynamic changes of hydrogen sulfide concentration in real time in a cell model, and its sensitivity reaches 0.1 μM, has good selectivity, and is not interfered by other biological molecules.

9. The use of a fluorescent probe for detecting hydrogen sulfide according to claim 7, characterized in that: The probe can be used for environmental monitoring, especially for real-time monitoring of hydrogen sulfide concentration in water quality monitoring, air quality assessment and soil pollution monitoring.

Images