Quinolyl quinazolinone-phenothiazine synergistic difunctional fluorescent probe compound as well as preparation method and application thereof

Abstract

The invention discloses a quinolyl quinazolinone-phenothiazine synergistic difunctional fluorescent probe compound as well as a preparation method and application thereof, and belongs to the technical field of fluorescent probes. According to the invention, a difunctional fluorescent probe (probe PT) with a multi-component synergistic effect is constructed, and high-selectivity and high-sensitivity visual detection of copper ions (Cu < 2 + >) and hypochlorite (ClO-) is realized. The probe PT has significant response to Cu < 2 + > within 10 s, the color of the solution is changed from yellow to purple, and the detection limit is 1.59 * 10 <-10 > mol / L; the response to ClO <-> shows that the solution becomes colorless from yellow, and the detection limit is as low as 4.3 * 10 <-9 > mol / L. Under the irradiation of a 365 nm ultraviolet lamp, the probe PT can realize the simultaneous fluorescence recognition of Cu < 2 + > and ClO <->. Besides, the probe PT is embedded in a hydrogel matrix to construct a portable detection plate, the colorimetric and fluorescence response of the portable detection plate are consistent with those of a solution system, a low-cost and feasible portable technical scheme is provided for on-site monitoring, prevention and control of Cu < 2 + > and ClO <-> pollution, and the portable detection plate has a good application prospect.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of fluorescent probes, and particularly relates to a quinolinyl quinazolinone-phenothiazine synergistic bifunctional fluorescent probe compound and a preparation method and application thereof. BACKGROUND

[0002] Copper ions (Cu 2+ ) and hypochlorite (ClO - ) are important inorganic substances widely used in industrial production, but improper use or discharge can cause serious harm to the environment and human health. Cu 2+ Participates in a variety of key metabolic processes in living organisms, is an important component of tyrosinase, monoamine oxidase, superoxide dismutase and plasma ceruloplasmin and other oxidation-related enzymes. Maintaining Cu 2+ Homeostasis is the basis for normal cell metabolism and organ function, while excessive Cu 2+ intake can cause diarrhea, vomiting, liver and kidney damage and other adverse reactions, and even be closely related to the occurrence of serious diseases such as Alzheimer's disease, Wilson's disease and Menkes disease. ClO - is a strong oxidizing substance, widely used as a bleaching agent and disinfectant in the food industry, wastewater treatment and medical and health fields, and is one of the indispensable chemicals in human daily life. At the same time, ClO - is also an important active oxygen species in the body, which plays a key role in immune defense, inflammatory response and pathogen clearance. However, abnormal elevation of ClO - level in the body can cause oxidative damage to tissues, and then induce atherosclerosis, kidney disease, ischemia-reperfusion injury, neurodegeneration, and even have a potential carcinogenic risk. In addition, ClO - is easy to enter the water cycle during use and discharge, causing drinking water and agricultural product pollution, and posing a threat to the ecological environment and human health. Therefore, developing a simple, sensitive and practical Cu 2+ and ClO - detection and analysis method is of great significance for water environment protection and human health safety.

[0003] In the practical application of Cu 2+ and ClO - detection, on-site rapid detection is an important problem to be solved. Traditional Cu 2+ and ClO -The analysis methods are mostly dependent on laboratory techniques such as chromatography and spectroscopy, which have high sensitivity and accuracy, but usually require complex sample pretreatment, bulky and expensive equipment and professional operators, and are difficult to meet the application requirements of on-site rapid and real-time detection. In contrast, portable detection technology gradually attracts attention due to its simple operation, easy portability and adaptability to on-site environment. Therefore, it is urgent to develop a portable detection method with reasonable structure design, rapid response, high sensitivity and visual recognition to meet the on-site rapid detection requirements of Cu 2+ and ClO - in complex environmental systems. SUMMARY

[0004] To overcome the deficiencies in the prior art, the present application is based on the metal-ligand charge transfer (MLCT) theory to design the molecular structure, taking quinoline and quinazolinone structural units as the recognition groups of Cu 2+ . At the same time, considering the luminescence performance of the probe and its recognition requirements for ClO - , a phenothiazine structural unit is introduced as a chromophore, which also has the function of recognizing ClO - . Through the synergistic integration of the above multifunctional structural units, a multi-component synergistic visual bifunctional fluorescent probe PT is synthesized. The probe PT shows good selectivity, rapid response characteristics and high detection sensitivity for Cu 2+ and ClO - , which provides a new technical idea for the design and synthesis of bifunctional fluorescent probes in the field of synergistic detection of Cu 2+ and ClO - .

[0005] To achieve the above technical problems, the present application adopts the following technical solutions: The purpose of the present application is to provide a quinoline-based quinazolinone-phenothiazine synergistic bifunctional fluorescent probe compound, and the structural formula of the fluorescent probe compound is as follows: .

[0006] The above fluorescent probe is obtained by the following synthesis steps, and the synthesis route is as follows:

[0007] Another purpose of the present application is to provide a preparation method of the above quinoline-based quinazolinone-phenothiazine synergistic bifunctional fluorescent probe compound.

[0008] A preparation method of a quinoline-based quinazolinone-phenothiazine synergistic bifunctional fluorescent probe compound, comprising the following steps: comprising the following steps: Step 1, 4-(10H-phenothiazin-10-yl)benzaldehyde, anthranilamide, copper chloride were added into anhydrous ethanol, and the reaction was carried out under reflux, and after cooling, the mixture was filtered, washed with anhydrous ethanol, and the intermediate was obtained; Step 2, the intermediate, 2-bromoquinoline, sodium tert-butoxide, [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride and 1,4-dioxane were mixed, and the reaction was carried out under heating and inert gas protection, and after the reaction was completed, the mixture was filtered, and then the filtrate was poured into ice water, and the solid was precipitated, and the fluorescent probe compound was obtained by purification.

[0009] Further limited, in step 1, 6.59 mmol of 4-(10H-phenothiazin-10-yl)benzaldehyde, 7.25 mmol of anthranilamide, and 659.24 μmol of copper chloride were added into 25 mL of anhydrous ethanol.

[0010] Further limited, in step 2, the inert gas is nitrogen or argon.

[0011] Further limited, in step 2, heating was carried out at 100-120°C.

[0012] Further limited, in step 2, column chromatography was used for purification, and the eluent used for purification was ethyl acetate and petroleum ether in a volume ratio of 1:10.

[0013] Further limited, in step 2, the reaction progress was monitored by thin layer chromatography (TLC).

[0014] In addition, the use of the quinolinyl quinazolinone-phenothiazine synergistic bifunctional fluorescent probe compound of the present application is also provided for the detection of Cu 2+ and ClO - .

[0015] The present application aims to provide a preparation method of a probe hydrogel detection plate, comprising the following steps: 1.0 g of PVA and 1.0 g of PEG were mixed with 20 mL of EtOH / H2O solution (1:1 v / v), and magnetically stirred at 90°C until completely dissolved to form a transparent viscous liquid, and a PVA / PEG hydrogel was obtained; 1.3 mg of the fluorescent probe compound of claim 1 or prepared by the method of any one of claims 2-8 was dissolved in 6 mL of DMF, and then added to the above PVA / PEG hydrogel, and mixed thoroughly to obtain a probe PT hydrogel; The probe PT hydrogel was taken with a rubber bulb dropper and dropped into a plastic mold, and cooled and shaped to obtain the detection plate.

[0016] Considering the on-site detection of Cu 2+ and ClO- The important parameters of the present application, the fluorescence spectrum titration method is further evaluated probe PT to Cu 2+ And ClO - The detection limit and response time. After adding copper ions, the emission intensity of probe PT decreases to the minimum value within 10 s, and there is a significant linear relationship between Cu 2+ Concentration, the linear regression equation is F 490 =27193.28994-480.71323x, the correlation coefficient R 2 =0.99128, the detection limit is calculated as 7.16×10 -10 mol / L. The detection of ClO - The probe PT can be reduced to the minimum value within 5 s, and the detection limit is as low as 4.3×10 -9 mol / L (linear regression equation F 490 =973.22727-91.87273x, the correlation coefficient R 2 =0.99554).

[0017] The present application also provides the application of the above-mentioned fluorescent probe. According to the visualization function of the fluorescent probe for Cu 2+ And ClO - Detection, a portable hydrogel detection plate based on probe PT is prepared, and its detection process is refined to effectively avoid interference and damage in the detection process. This provides a low-cost, rapid and sensitive portable detection method for Cu 2+ And ClO - .

[0018] Compared with the prior art, the present application has the following beneficial effects: The present application combines quinoline-quinazolinone structural unit with phenothiazine structural unit, and innovatively constructs two multifunctional fluorescent probes PT with multi-component synergistic effect. The probe PT can realize high selectivity, high sensitivity and visual detection of Cu 2+ And ClO - , and has the key performance parameters required for on-site rapid detection. In terms of detection performance, the response of probe PT to Cu 2+ Can be completed within 10 s, and the solution color changes from yellow to purple, and the minimum detection limit reaches 1.59×10 - 10 mol / L; the response to ClO - The solution color changes from yellow to colorless, and the detection limit is as low as 4.3×10 -9 mol / L. The probe PT to Cu 2+ And ClO -exhibited excellent sensitivity and selectivity, and under the irradiation of 365 nm ultraviolet light, the specific detection of Cu 2+ and ClO - can be realized.

[0019] The present application also successfully embeds the probe PT in the hydrogel matrix, constructs a portable hydrogel detection plate, and verifies its actual application performance, which shows that the hydrogel detection plate is consistent with the probe PT solution system in terms of colorimetric and fluorescent response, and provides a portable, low-cost and simple operation detection scheme for the on-site monitoring of Cu 2+ and ClO - , which has good application potential and promotional value.

[0020] For a better understanding of the features and technical contents of the present application, please refer to the detailed description and the accompanying drawings. It should be noted that the accompanying drawings are provided for illustrative purposes only and are not intended to limit the present application. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 is the H-NMR spectrum of intermediate 2 1 . Figure 2 is the H-NMR spectrum of probe PT 1 . Figure 3 is the C-NMR spectrum of probe PT 13 . Figure 4 is the specific detection of Cu 2+ and ClO - by probe PT.

[0022] Figure 5 is the preparation and detection flowchart of the hydrogel detection plate. Figure 6 is the detection ability of the hydrogel detection plate based on probe PT for Cu 2+ , ClO - and Cu 2+ + ClO - . DETAILED DESCRIPTION

[0023] The present application will be described in detail below in combination with specific examples. These examples are helpful for those skilled in the art to further understand the present application, but should not be regarded as limiting the present application. It should be noted that for those skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made. These all belong to the protection scope of the present application.

[0024] Example 1: Synthesis of intermediate 2 Into a 50 mL round bottom flask, 4-(10H-phenothiazin-10-yl)benzaldehyde (2.00 g, 6.59 mmol), anthranilic acid (987.34 mg, 7.25 mmol), copper chloride (88.63 mg, 659.24 pmol) and 25 mL of anhydrous ethanol were added. The mixture was refluxed for 5 h, and after cooling, the solid was collected by filtration. The product was washed with ethanol to obtain intermediate 2, yellow-green solid 3.0 g, yield 92%. 1 H NMR (400 MHz, DMSO) δ 12.79 (s, 1H), 8.57 (d, 2H), 8.40 (d,2H), 8.21 (d, 1H), 7.89 (t, 1H), 7.79 (dd, 3H), 7.67 (t, 2H), 7.58 (t, 1H),7.36 (t, 2H), 6.79 (d, 2H), spectrum as shown in Figure 1 .

[0025] The structural formula of intermediate 2 is as follows:

[0026] Example 2: Synthesis of target compound probe PT Into a dry 50 mL reaction flask, intermediate 2 (1.00 g, 2.38 mmol), 2-bromoquinoline (545.56 mg, 2.62 mmol), sodium tert-butoxide (297.82 mg, 3.10 mmol), [1,1’- bis(diphenylphosphino)ferrocene]palladium(II) dichloride (174.42 mg, 238.38 pmol) and 15 mL of 1,4-dioxane were added. The reaction was carried out under nitrogen protection at 110°C, and the progress of the reaction was monitored by thin layer chromatography (TLC). After the reaction was completed, the reaction solution was filtered, and the filtrate was poured into ice water to precipitate a brown-yellow solid. The crude product was purified by column chromatography (eluent: ethyl acetate: petroleum ether = 1:10) to obtain the target product probe PT, white solid 1.03 g, yield 79%. 1 H NMR (400 MHz, Chloroform- d ) δ 8.85 (d, 3H), 8.29 (dd, 7H), 7.94 - 7.71 (m, 8H), 7.58 (t, 4H), spectrum as shown in Figure 2 . 13 C-NMR (101 MHz, Chloroform- d) δ 155.2, 146.9, 135.8, 128.9, 128.6, 127.4, 126.7, 126.0, 118.4, spectrum as shown in Figure 3 .

[0027] Probe PT for Cu 2+ and ClO - bifunctional recognition The present application respectively verifies the detection specificity of probe PT for Cu 2+ and ClO - , as shown in Figure 4 . Hg 2+ , Mg 2+ , Al 3+ , Cu 2+ , Pb 2+ , Mn 2+ , Zn 2+ , Ag + , Fe 3+ , Ni 2+ , Cu + and Na + and other metal cations are selected as interference for investigation. When the above interference (concentration is 60 μM) is added to the solution of probe PT, the fluorescence intensity does not change obviously. However, after adding 60 μM of Cu 2+ , the fluorescence intensity of probe PT is significantly reduced, and the color of the solution can be observed to change from yellow to purple under sunlight. Further, different kinds of anions (ClO - , Cl - , Br - , SO4 2- , CO3 2- , OH - , HCO3 - , SO3 2- , S2O3 2- ) are added to the solution of probe PT for testing, and the results show that only when ClO - exists, the solution can change from yellow to colorless obviously, and under the irradiation of ultraviolet lamp (365 nm), the blue fluorescence intensity in the solution with ClO - exists is significantly reduced, indicating that probe PT has good selectivity and visual response ability for ClO - . In addition, the present application further investigates the recognition performance of probe PT for ClO 2 + under the coexistence of Cu - . 2+ - ​In the mixed system, the probe PT solution can still change from yellow to colorless. Under the irradiation of 365 nm ultraviolet light, the fluorescence of the solution is obviously weakened, and yellow-green emission is presented, indicating that the probe can realize the detection of Cu 2+ and ClO - at the same time.

[0028] Preparation of the probe PT hydrogel detection plate Firstly, 1.0 g of PVA and 1.0 g of PEG are mixed with 20 mL of EtOH / H2O solution (1:1 v / v), and are magnetically stirred at 90 DEG C until completely dissolved to form a transparent viscous liquid.

[0029] Then, the probe PT hydrogel is prepared: 1.3 mg of the probe PT is dissolved in 6 mL of DMF, and then is added into the above PVA / PEG hydrogel and mixed thoroughly.

[0030] Finally, the preparation of the hydrogel detection plate is prepared: the probe PT hydrogel is taken by using a rubber head dropper, and is dropped into a plastic mold and cooled to shape.

[0031] The application adds the use of a sampling tube in the detection process, the cover of the sampling tube is provided with an inserted sealing inner plug, the top of which is integrally formed with a droplet opening, which is used for extruding liquid for detection. In order to avoid the interference of solid impurities during on-site detection, a small group of cotton is inserted into the droplet opening to play a simple filtering role, and the specific preparation and detection process is shown in Figure 5 .

[0032] Detection effect of the hydrogel detection plate The application embeds the probe PT in the hydrogel matrix to prepare a hydrogel detection plate, and detects under the conditions of blank and adding ClO - , Cu 2+ or Cu 2+ / ClO - coexistence, as shown in Figure 6 . The blank detection plate shows a strong blue fluorescence under natural light, and the color changes from yellow to colorless after adding ClO - , and the fluorescence is obviously weakened; after adding Cu 2+ , the color is obviously changed to purple, and the blue fluorescence is almost completely quenched under the coexistence of Cu 2+ and ClO - , the fluorescence is further weakened and presents different fluorescence characteristics from the single ion, indicating that the probe PT can realize the effective differentiation and simultaneous detection of Cu 2+ and ClO - in the hydrogel matrix, and the colorimetric and fluorescence response behaviors are consistent with the solution system, which verifies the feasibility and practicability of the hydrogel detection plate in on-site rapid detection.

[0033] The specific embodiments of the present application are described above. It should be noted that the present application is not limited to the specific embodiments described above. Various modifications and changes can be made without departing from the scope of the present application as defined in the claims.

Claims

1. A quinolinylquinazolinone-phenothiazine synergistic bifunctional fluorescent probe compound, characterized in that, The fluorescent probe has the following structural formula: 。 2. The method for preparing the fluorescent probe compound according to claim 1, characterized in that, The method comprises the following steps: Step 1: 4-(10H-phenothiazine-10-yl) benzaldehyde, o-aminobenzamide and copper chloride are added into anhydrous ethanol, and refluxed, and then filtered after cooling, washed with anhydrous ethanol to obtain an intermediate; Step 2: the intermediate, 2-bromoquinoline, sodium tert-butoxide, [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) dichloride and 1,4-dioxane are mixed, and heated under inert gas protection, and then filtered after the reaction is completed, and then the filtrate is poured into ice water to precipitate a solid, and then purified to obtain the fluorescent probe compound.

3. The method of claim 2, wherein, In step 1, 6.59 mmol of 4-(10H-phenothiazine-10-yl) benzaldehyde, 7.25 mmol of o-aminobenzamide and 659.24 μmol of copper chloride are added into 25 mL of anhydrous ethanol.

4. The method of claim 2, wherein, In step 2, the inert gas is nitrogen or argon.

5. The method of claim 2, wherein, In step 2, heating is performed at 100-120°C.

6. The method of claim 2, wherein, In step 2, column chromatography is used for purification.

7. The method of claim 6, wherein, The eluent used for purification is ethyl acetate and petroleum ether in a volume ratio of 1:

10.

8. The method of claim 3, wherein, In step 2, thin layer chromatography (TLC) is used to monitor the progress of the reaction.

9. The fluorescent probe compound as claimed in claim 1 for the detection of Cu 2+ and CIO - and CIO 10. A method for preparing a probe hydrogel detection plate, characterized in that, The method comprises the following steps: 1.0 g of PVA and 1.0 g of PEG are mixed with 20 mL of an EtOH / H2O solution (1:1 v / v) and magnetically stirred at 90°C until completely dissolved to form a transparent viscous liquid, thereby obtaining a PVA / PEG hydrogel; 1.3 mg of the fluorescent probe compound of claim 1 or prepared by the method of any one of claims 2-8 is dissolved in 6 mL of DMF, and then added into the PVA / PEG hydrogel, and mixed thoroughly, thereby obtaining a probe PT hydrogel; The probe PT hydrogel is taken by using a rubber bulb dropper, and then dropped into a plastic mold, and then cooled and shaped, thereby obtaining the detection plate.

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