Preparation and application of a fluorescent probe for detecting aluminum ions

Abstract

The application discloses a kind of super-sensitive aluminum ion detection fluorescent probe preparation and application, it is related to converter station valve internal cooling water system aluminum ion fluorescent probe technical field.Probe structure formula is: preparation method is: 4-amino benzhydrazide and 2-hydroxy-1-naphthaldehyde are dissolved in anhydrous ethanol, then reflux, product solution is filtered under vacuum to obtain filtrate, and under the condition of reduced pressure, rotary evaporation dry solvent, obtain benzhydrazide compound based on Schiff base functional group.The application of probe is used to detect the concentration of aluminum ion in sample.The aluminum ion fluorescent probe of the application can be reacted with aluminum ion, and the change of fluorescence spectrum is generated, so that the quantitative detection of aluminum ion is realized.The probe of the application is very sensitive to aluminum ion reaction, so as to facilitate the rapid detection of aluminum ion.The stability of fluorescent probe is good, and then it can be stored for a long time.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of aluminum ion fluorescent probe technology of valve inner cooling water system of converter station, and particularly relates to a super-sensitive aluminum ion detection fluorescent probe preparation and application. BACKGROUND

[0002] The converter valve can realize AC-DC conversion and plays an important role in the extra-high voltage direct current transmission system. The aluminum radiator is generally used in the inner cooling water system of the converter valve, and the electrochemical reaction of the aluminum radiator in water is the root cause of the corrosion and fouling of the inner cooling water system. According to the DL / T 1716-2017 'Guidelines for Operation and Management of Cooling Water of High Voltage Direct Current Transmission Converter Valve', the aluminum ion concentration in the inner cooling water should be controlled below 2 μg / L. Therefore, it is urgent to develop an analysis technology with high selectivity, high sensitivity, rapidity, convenience and non-destructiveness to detect the content of aluminum ions in the inner cooling water, so as to detect the corrosion of the aluminum radiator in time and ensure the safe operation of the converter valve.

[0003] In recent years, ion chromatography (IC), atomic absorption spectrometry (AAS), inductively coupled plasma mass spectrometry (ICP-MS) and fluorescent probe analysis method have been used for the determination and analysis of various substances. The fluorescent probe analysis method has attracted widespread attention due to its high sensitivity, good selectivity, rapid response, simple operation, non-destructive detection and excellent imaging quality. This method can accurately and quantitatively analyze target substances under complex conditions. However, most of the reported fluorescent probes are plagued by problems such as high synthesis cost, complex synthesis, low sensitivity, poor selectivity and poor water solubility. In addition, other metal ions existing in the environment and in the body may potentially interfere with the detection of aluminum ions.

[0004] In summary, it is urgent to develop a fluorescent probe with high sensitivity, simple synthesis, good water solubility, rapid response and specific recognition, which will help to overcome the limitations of current technology and improve the detection accuracy of aluminum ions, and meet the needs of the aluminum ion detection field in the inner cooling water of the converter valve. SUMMARY

[0005] To solve the above technical problems, the present application discloses a super-sensitive aluminum ion detection fluorescent probe preparation and application. The aluminum ion fluorescent probe of the present application has simple synthesis, high sensitivity, rapid response and can specifically recognize aluminum ions.

[0006] To achieve the above purpose, the present application adopts the following technical solutions:

[0007] The present application discloses a benzoyl hydrazine compound based on a Schiff base functional group, and the structural formula is as follows:

[0008]

[0009] wherein R1, R2, R3, R4, R5, R6, R7, R8, R9 and R 10 are free hydrogen atoms, straight-chain or branched alkyl groups, straight-chain or branched alkoxy groups, sulfonic acid groups, ester groups, carboxyl groups; R1, R2, R3, R4, R5, R6, R7, R8, R9 and R 10 may be the same or different.

[0010] The second aspect of the present application provides a preparation method of the above-mentioned benzhydrazide compound based on the Schiff base functional group, and the synthesis route is as follows:

[0011]

[0012] The benzhydrazide compound and the 2-hydroxy-1-naphthaldehyde compound are dissolved in anhydrous ethanol, refluxed, the product solution is vacuum filtered to obtain a filtrate, and the solvent is dried by rotary evaporation under reduced pressure to obtain the benzhydrazide compound based on the Schiff base functional group.

[0013] Optionally, a benzhydrazide compound based on the Schiff base functional group has the following structural formula:

[0014]

[0015] that is, in the above-mentioned structural formula, R1, R2, R3, R4, R5, R6, R7, R8, R9 and R 10 are all free hydrogen atoms.

[0016] The third aspect of the present application provides a super-sensitive aluminum ion detection fluorescent probe, which comprises the above-mentioned benzhydrazide compound based on the Schiff base functional group, and has the following structural formula:

[0017]

[0018] The fourth aspect of the present application provides a preparation method of the above-mentioned super-sensitive aluminum ion detection fluorescent probe, which comprises the following steps:

[0019] The 4-aminobenzhydrazide and the 2-hydroxy-1-naphthaldehyde are dissolved in anhydrous ethanol, then refluxed, the product solution is vacuum filtered to obtain a filtrate, and the solvent is dried by rotary evaporation under reduced pressure to obtain the benzhydrazide compound based on the Schiff base functional group.

[0020] Optionally, the molar ratio of the 4-aminobenzhydrazide and the 2-hydroxy-1-naphthaldehyde is 1:1-3:1.

[0021] Optionally, the reflux time is 2-8 h.

[0022] Optionally, the pressure of the reduced pressure condition is-0.08 to-0.1 Mpa.

[0023] Optionally, the benzhydrazide compound based on the Schiff base functional group is further purified by passing through a liquid chromatography column using a mixture of dichloromethane and methanol (v:v = 30:1).

[0024] The fifth aspect of the present application provides an application of the above-mentioned super-sensitive aluminum ion detection fluorescent probe, which is applied to detecting the aluminum ion concentration in a sample.

[0025] Optionally, the application comprises the step of contacting the super-sensitive aluminum ion detection fluorescent probe with the sample.

[0026] Optionally, the sample is water or a biological cell.

[0027] The sixth aspect of the present application provides a detection preparation comprising the above-mentioned super-sensitive aluminum ion detection fluorescent probe.

[0028] Optionally, the detection preparation further comprises a buffer for determining the aluminum ion concentration in the sample.

[0029] Optionally, the detection preparation further comprises a user manual.

[0030] The seventh aspect of the present application provides an application of the above-mentioned detection preparation, which is applied to determining the aluminum ion concentration in a sample.

[0031] Optionally, the sample is water or a biological cell.

[0032] The present application has the following beneficial effects,

[0033] 1. The aluminum ion fluorescent probe of the present application can react with aluminum ions to produce changes in fluorescence spectrum, thereby realizing quantitative detection of aluminum ions. Specifically, the aluminum ion fluorescent probe of the present application cannot react with zinc ions, magnesium ions, calcium ions, trivalent iron ions, divalent iron ions, potassium ions, sodium ions, chloride ions, sulfate ions, etc. commonly found in cold water to cause significant changes in fluorescence spectrum, thereby realizing highly selective recognition of aluminum ions.

[0034] 2. The aluminum ion fluorescent probe of the present application is very sensitive to aluminum ions, thereby facilitating rapid detection of aluminum ions.

[0035] 3. The aluminum ion fluorescent probe of the present application has good stability and can be stored for a long time.

[0036] In summary, the aluminum ion fluorescent probe of the present application is a highly selective super-sensitive aluminum ion fluorescent probe, which is simple to synthesize and is conducive to commercialization and application. BRIEF DESCRIPTION OF DRAWINGS

[0037] Figure 1 (a) is the fluorescence spectrum before and after the probe (10 μM) is added to aluminum ions (0-20 μM); Figure 1(b) is the working curve of quantitative analysis of aluminum ions (0-20 μM) at different concentrations using a probe (10 μM); the excitation wavelength is 395 nm, and both the excitation and emission slits are 5 nm.

[0038] Figure 2 This shows the change in the spectrum over time after the addition of aluminum ions (20 μM) to the probe (10 μM);

[0039] Figure 3 This shows the effects of common ions, reactive oxygen species, and amino acids on the fluorescence intensity of the probe (10 μM) and the response of the probe to aluminum ions in the presence of different analytes. The bar chart represents the fluorescence intensity of the probe at 475 nm in the presence of different analytes.

[0040] Figure 4 The effects of different pH values ​​on the probe (10 μM) and the response of the probe (10 μM) to aluminum ions (20 μM) under different pH conditions;

[0041] Figure 5 This is an analysis of the toxicity of various probe concentrations to HeLa cells, with concentrations of 0 μM, 5 μM, and 10 μM.

[0042] Figure 6 This describes the recognition performance of a 10 μM probe on aluminum ions in HeLa cells. A: Blank control group cells; B: Cells incubated with the 10 μM probe for 20 minutes; C: Cells incubated with the 10 μM probe for 20 minutes, followed by incubation with 20 μM aluminum ions for 20 minutes; D: Cells incubated with the 10 μM probe for 20 minutes, followed by incubation with 50 μM aluminum ions for 20 minutes; E: Cells incubated with the 10 μM probe for 20 minutes, followed by incubation with 100 μM aluminum ions for 20 minutes.

[0043] F: Bar graph of cell fluorescence intensity, scale bar 20 μm, excitation wavelength 395 nm, collected emission wavelength 430-530 nm.

[0044] nm;

[0045] Figure 7 The results show the recognition performance of the probe (10 μM) on aluminum ions in zebrafish. A: blank control group zebrafish; B: zebrafish incubated with the probe for 20 minutes; C: zebrafish incubated with the probe (10 μM) for 20 minutes, then incubated with aluminum ions (50 μM) for 20 minutes; D: bar graph of AC fluorescence intensity; scale bar is 100 μm, excitation wavelength is 395 nm, and the collected emission wavelength is 430-530 nm. Detailed Implementation

[0046] With reference to the accompanying drawings, the technical solutions in the embodiments of the present application will be clearly and completely described below, obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work belong to the protection scope of the present application.

[0047] Embodiment 1

[0048] A benzoyl hydrazine compound based on a Schiff base functional group, a structural formula of which is as follows:

[0049]

[0050] In the formula, R1, R2, R3, R4, R5, R6, R7, R8, R9 and R 10 are free hydrogen atoms, straight-chain or branched alkyl groups, straight-chain or branched alkoxy groups, sulfonic acid groups, ester groups, carboxyl groups; R1, R2, R3, R4, R5, R6, R7, R8, R9 and R 10 may be the same or different.

[0051] The preparation method of the above-mentioned benzoyl hydrazine compound based on a Schiff base functional group is as follows:

[0052]

[0053] Alternatively, a benzoyl hydrazine compound based on a Schiff base functional group, a structural formula of which is as follows:

[0054]

[0055] In the above-mentioned structural formula, R1, R2, R3, R4, R5, R6, R7, R8, R9 and R 10 are all free hydrogen atoms.

[0056] Embodiment 2

[0057] A super-sensitive aluminum ion detection fluorescent probe, comprising the benzoyl hydrazine compound based on a Schiff base functional group described in Embodiment 1, a structural formula of which is as follows:

[0058]

[0059] The preparation method of the above-mentioned super-sensitive aluminum ion detection fluorescent probe comprises the following steps:

[0060] 4-Aminobenzoyl hydrazine and 2-hydroxy-1-naphthaldehyde are dissolved in anhydrous ethanol, then refluxed, the product solution is vacuum filtered to obtain a filtrate, and the solvent is rotary evaporated under reduced pressure to obtain the benzoyl hydrazine compound based on a Schiff base functional group.

[0061] Optionally, the molar ratio of the 4-aminobenzhydrazide and 2-hydroxy-1-naphthaldehyde is 1:1-3:1.

[0062] Optionally, the refluxing time is 2-8h.

[0063] Optionally, the pressure of the reduced pressure condition is -0.08--0.1Mpa.

[0064] Optionally, the method further comprises purifying the benzhydrazide compound based on the Schiff base functional group by passing it through a liquid chromatography column using a mixture of dichloromethane and methanol (v:v=30:1).

[0065] In the above preparation method, the amount of anhydrous ethanol is not limited, as long as it can dissolve the 4-aminobenzhydrazide and 2-hydroxy-1-naphthaldehyde. In addition, the specific parameters for vacuum filtration are not limited, and conventional operations are used or adjusted according to specific conditions.

[0066] The above super-sensitive aluminum ion detection fluorescent probe is applied to detect the concentration of aluminum ions in a sample.

[0067] Optionally, the application comprises the step of contacting the super-sensitive aluminum ion detection fluorescent probe with the sample to be detected.

[0068] Optionally, the sample is water or a biological cell.

[0069] Example 3

[0070] A detection preparation comprising the above super-sensitive aluminum ion detection fluorescent probe.

[0071] Optionally, the detection preparation further comprises a buffer for determining the concentration of aluminum ions in the sample.

[0072] Optionally, the detection preparation further comprises instructions for use.

[0073] The above detection preparation is applied to determine the concentration of aluminum ions in a sample.

[0074] Optionally, the sample is water or a biological cell.

[0075] Application Example 1

[0076] 453mg (0.3mmol) of 4-aminobenzhydrazide was dissolved in 20mL of anhydrous ethanol, and then 688mg (0.4mmol) of 2-hydroxy-1-naphthaldehyde was added and heated to reflux for 4h. The resulting product solution was vacuum filtered to obtain a filtrate. To speed up the evaporation rate, the solvent was rotary evaporated under reduced pressure (pressure range: -0.08--0.1Mpa) to obtain a crude product. The crude product was passed through a liquid chromatography column using a mixture of dichloromethane and methanol (v:v=30:1) to obtain 705mg of yellow pure product, with a yield of 77%.

[0077]

[0078] A plurality of parallel samples with a probe concentration of 10 μM were configured in 10 mL cuvettes, and then different concentrations of aluminum ions were added to the test system, which was shaken uniformly and then left to stand. The above determination was performed in a (10 mM HEPES, pH 7.4) system, the probe used was the probe prepared in Application Example 1, and all spectral tests were measured at 25°C.

[0079] The fluorescence intensity change was tested by a fluorescence spectrometer, from which Figure 1 (a) It can be clearly seen that the fluorescence intensity at 475 nm gradually increased with the increase of the concentration of aluminum ions. Moreover, the fluorescence intensity of the probe (10 μM) added with aluminum ions (0-20 μM) showed a good linear relationship with the concentration of aluminum ions, which proved that the fluorescence probe could be used for quantitative analysis of aluminum ions. Figure 1 (b) It can be seen that the fluorescence intensity of the probe (10 μM) added with aluminum ions (0-20 μM) showed a good linear relationship with the concentration of aluminum ions, which proved that the fluorescence probe could be used for quantitative analysis of aluminum ions.

[0080] Figure 2 The fluorescence intensity change was tested by a fluorescence spectrometer, from which

[0081] It can be clearly seen that the fluorescence intensity reached a maximum value within 3 min after the addition of aluminum ions and remained unchanged, which indicated that the probe reacted rapidly with aluminum ions and could provide a rapid analysis method for the determination of aluminum ions. Figure 2

[0082] Figure 3 ​The figure is the influence of different analytes on the fluorescence spectrum of the probe (10 μM). The analytes are blank, zinc ion, tin ion, lead ion, nickel ion, manganese ion, magnesium ion, calcium ion, ferric ion, ferrous ion, potassium ion, sodium ion, silver ion, cerium ion, mercury ion, chloride ion, sulfate ion, carbonate ion, nitrate ion, hypochlorite, hydrogen peroxide, and cysteine (the concentration of other analytes is 100 μM except for special indication). The histogram represents the fluorescence intensity value of the probe at 475 nm in the presence of different analytes. The above determination is carried out in pure water (10 mM HEPES, pH 7.4), the probe used is the probe prepared in the application example 1, and all the spectrum tests are measured at 25 °C. Specifically, a plurality of parallel samples of the probe with a concentration of 10 μM are configured in 10 mL cuvettes, then a certain amount of analyte is added, shaken, and determined after 3 minutes.

[0083] From Figure 3 It can be clearly seen that only the addition of aluminum ion can cause a strong change in the fluorescence intensity of the probe, and the influence of other analytes can be almost ignored and does not significantly interfere with the fluorescence intensity of the probe for aluminum ion detection. The experiment proves that the probe has high selectivity for aluminum ion, which is conducive to the detection and analysis of aluminum ion.

[0084] Figure 4 The figure is the influence of different pH on the probe (10 μM) and the response of the probe (10 μM) to aluminum ion (20 μM) under different pH conditions. The above determination is carried out in pure water (10 mM HEPES, pH 7.4), the probe used is the probe prepared in the application example 1, and all the spectrum tests are measured at 25 °C.

[0085] From Figure 4 It can be clearly seen that the probe (10 μM) can exist stably in a wide pH range and respond to aluminum ion, and the response effect is best under neutral conditions.

[0086] Figure 5 The figure is the toxicity analysis of each probe concentration on HeLa cells. The probe concentrations are 0 μM, 5 μM, and 10 μM. The cell counting kit (CCK-8) is used to detect the cytotoxicity of different concentrations of the probe on HeLa cells, and the incubation time of the probe on the cells is 2 h. The probe used is the probe prepared in the application example 1.

[0087] From Figure 5 It can be clearly seen that the probe has the characteristics of low toxicity, and can be used for real-time detection of aluminum ion in cell samples for a long time.

[0088] Figure 6Fig. 6 is the recognition performance of the probe (10 μM) to aluminum ions in HeLa cells. A: cells incubated with the probe (10 μM) for 20 min; B: cells incubated with the probe (10 μM) for 20 min and then added with aluminum ions (20 μM) for 20 min; C: cells incubated with the probe (10 μM) for 20 min and then added with aluminum ions (50 μM) for 20 min; D: cells incubated with the probe (10 μM) for 20 min and then added with aluminum ions (100 μM) for 20 min. The probe used is the one prepared in Application Example 1, and the test is measured at 25 °C. The scale is 20 μm, the excitation wavelength is 395 nm, and the collected emission wavelength is 430-530 nm.

[0089] It can be clearly seen that the probe can detect aluminum ions in HeLa cells Figure 6

[0090] Figure 7 Fig. 7 is the recognition performance of the probe (10 μM) to aluminum ions in zebrafish. A: blank control group of zebrafish; B: zebrafish incubated with the probe for 20 min; C: zebrafish incubated with the probe (10 μM) for 20 min and then added with aluminum ions (50 μM) for 20 min; D: columnar chart of the fluorescence intensity of A-C. The probe used is the one prepared in Application Example 1, and the test is measured at 25 °C. The scale is 100 μm, the excitation wavelength is 395 nm, and the collected emission wavelength is 430-530 nm.

[0091] It can be clearly seen that the probe can image aluminum ions in zebrafish. The experiment proves that the probe can be applied to the detection of aluminum ions in biological samples. Figure 7 Application Example 2

[0092] The 453 mg (0.3 mmol) of 4-aminobenzhydrazide was dissolved in 20 mL of anhydrous ethanol, and then 172 mg (0.1 mmol) of 2-hydroxy-1-naphthaldehyde was added and heated to reflux for 4 h. The obtained product solution was vacuum filtered to obtain a filtrate. In order to accelerate the evaporation speed, the solvent was evaporated under reduced pressure (pressure range: -0.08 to -0.1 Mpa) to obtain a crude product. The crude product was subjected to liquid chromatography column using a mixture of dichloromethane and methanol (v:v=30:1) to obtain 448.35 mg of yellow pure product, with a yield of 49%.

[0093] Application Example 3

[0094]

[0095] ​​To 453 mg (0.3 mmol) of 4-aminobenzohydrazide dissolved in 20 mL of absolute ethanol, 516 mg (0.3 mmol) of 2-hydroxy-l-naphtaldehyde was added and heated to reflux for 4 h. The resulting product solution was vacuum filtered to obtain a filtrate. To speed up the evaporation, the solvent was evaporated under reduced pressure (pressure range: -0.08 to -0.1 MPa). The crude product was purified by liquid chromatography using a mixture of dichloromethane and methanol (v:v = 30:1) to obtain 751 mg of a yellow pure product with a yield of 82%.

[0096] Application Example 4

[0097] To 453 mg (0.3 mmol) of 4-aminobenzohydrazide dissolved in 20 mL of absolute ethanol, 516 mg (0.3 mmol) of 2-hydroxy-l-naphtaldehyde was added and heated to reflux for 4 h. The resulting product solution was vacuum filtered to obtain a filtrate. To speed up the evaporation, the solvent was evaporated under reduced pressure (pressure range: -0.08 to -0.1 MPa). The crude product was purified by liquid chromatography using a mixture of dichloromethane and methanol (v:v = 30:1) to obtain 751 mg of a yellow pure product with a yield of 82%.

[0098] Application Example 5

[0099] To 453 mg (0.3 mmol) of 4-aminobenzohydrazide dissolved in 20 mL of absolute ethanol, 516 mg (0.3 mmol) of 2-hydroxy-l-naphtaldehyde was added and heated to reflux for 4 h. The resulting product solution was vacuum filtered to obtain a filtrate. To speed up the evaporation, the solvent was evaporated under reduced pressure (pressure range: -0.08 to -0.1 MPa). The crude product was purified by liquid chromatography using a mixture of dichloromethane and methanol (v:v = 30:1) to obtain 751 mg of a yellow pure product with a yield of 82%.

[0100] Of course, the above description is not a limitation of the present application, and the present application is not limited to the above examples. Changes, modifications, additions or substitutions made by those skilled in the art within the scope of the present application should also be included in the scope of the present application.

Claims

1. Use of an aluminum ion-detecting fluorescent probe, characterized in that, The application is applied to detecting the concentration of aluminum ions in a sample; The method comprises the step of contacting the aluminum ion detection fluorescent probe with the sample; The sample is water or biological cells; The aluminum ion detection fluorescent probe comprises a benzhydrazide compound based on a Schiff base functional group, and has the following structural formula:

2. A test for the use of a preparation, characterized in that The application is applied to detecting the concentration of aluminum ions in a sample; The sample is water or biological cells; The detection reagent comprises an aluminum ion detection fluorescent probe, and the aluminum ion detection fluorescent probe is a benzhydrazide compound based on a Schiff base functional group, and has the following structural formula:

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