Application of phenazine derivative in detection of p-phenylenediamine
The phenazine derivative DCP is used as a fluorescence sensor to solve the problem of complex and inconvenient detection of p-phenylenediamine, real-time detection with high selectivity and high sensitivity is achieved, and it is suitable for portable detection devices.
Patent Information
- Application Number
- CN202510347102.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-29
- Publication Date
- 2025-08-22
AI Technical Summary
The detection methods of p-phenylenediamine (p-PD) in the prior art are complex and inconvenient to carry, and real-time detection cannot be achieved. The sensor development is less and difficult to operate.
The phenazine derivative DCP was used as a fluorescence sensor to prepare the phenazine derivative DCP through the synthetic route by reacting ortho-phenylenediamine and 2,5-dihydroxy-1,4-benzoquinone and other compounds, which were used for the high-selective fluorescence identification and detection of phenylene diamine, and combined with the detection test strips to achieve portable detection.
It realizes high sensitivity, low detection limit (0.396 nM) and high selectivity fluorescence detection to p-PD, and can detect p-phenylenediamine in real time and conveniently, with good anti-interference ability.
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Abstract
Description
Technical Field
[0001] The invention relates to application of a phenazine derivative in detecting p-phenylenediamine, and belongs to the field of isomer detection. Background Art
[0002] Paraphenylenediamine (p-PD) is one of the simplest aromatic amines and is slightly soluble in cold water and most organic solvents. p-PD possesses all the properties of an aromatic ring and an amino group. It is widely used in the manufacture of azo, sulfide, and fur dyes, as well as rubber antioxidants and synthetic resins. p-PD is primarily used in black or dark hair dyes to achieve a long-lasting color. p-PD can be absorbed through the respiratory tract and skin. Although the percutaneous absorption rate is not very high, topical application or ingestion may cause local or systemic toxicity. p-PD can cause allergic dermatitis and acute renal failure and is a suspected carcinogen for bladder and hematopoietic tumors.
[0003] To date, very few sensors have been developed to detect p-PDs. These sensors are not portable, making them difficult to detect in real time. Existing detection methods primarily rely on fluorescence detectors, making them impractical for everyday use and complex to operate. Therefore, further sensor research needs to address these issues to achieve real-time detection of p-PDs.
[0004] Phenazines are organic natural products primarily used in dyes, pharmaceuticals, organic synthesis intermediates, and biochemical research. They possess an electron-deficient π system, a pair of electron-bearing nitrogen atoms, and three fused aromatic rings. They possess structural tunability, excellent optical properties, and high quantum yields, making them suitable for use as ionic ligands and hydrogen bond acceptors. They also favor π-π electron overlap, making them promising compounds for supramolecular chemistry and a wide range of applications in sensors. Building on our previous research, we investigated the use of phenazine derivatives as p-phenylenediamine (p-PD) fluorescence sensors (DCPs). These DCPs exhibit excellent p-PD sensing performance, including exceptional sensitivity, high specificity, instantaneous response, and good biocompatibility. They can selectively recognize p-PDs through fluorescence, exhibiting specific selectivity, low detection limits, and minimal environmental impact. Summary of the Invention
[0005] The purpose of the present invention is to provide the use of phenazine derivatives in detecting p-PD.
[0006] 1. Synthesis of phenazine derivative DCP Phenazine derivative DCP, whose molecular formula is C 16 H 12 N2O6, the structural formula is: The synthesis method of the phenazine derivative sensing molecule DCP comprises the following steps: (1) Synthesis of intermediate compound 1: Using water as solvent, o-phenylenediamine and 2,5-dihydroxy-1,4-benzoquinone were refluxed at 100°C in a molar ratio of 1:1 to 1:1.5 for 24 h, and filtered under reduced pressure. The solid was dissolved in a hot alkaline solution, filtered while hot, and the pH of the solution was adjusted to 3-4. The solution was cooled and allowed to stand, and filtered under reduced pressure to obtain intermediate compound 1.
[0007] (2) Synthesis of intermediate compound 2: Using acetonitrile as solvent and potassium carbonate and benzyltriethylammonium chloride as co-catalysts, intermediate compound 1 and ethyl bromoacetate were reacted at a molar ratio of 1:2.2~2.5 under nitrogen protection at 85°C for 15h. The mixture was filtered while hot to remove inorganic salts, extracted with dichloromethane and distilled water, dried over anhydrous sodium sulfate, and the organic phase was separated by column chromatography to obtain a light yellow solid, which was intermediate compound 2. (3) Synthesis of compound DCP: Using DMF as solvent, the intermediate compound 2 and sodium hydroxide (NaOH) were reacted at a molar ratio of 1:5-6 at 120°C for 20 h, acidified to 2-3 with hydrochloric acid, filtered under reduced pressure, and recrystallized from ethanol to obtain the final compound DCP. The synthetic route is as follows: .
[0008] 2. Detection of DCP by Phenazine Derivatives 1. Fluorescence recognition performance of p-PD The concentration was 2.0×10 -5 To a 10-fold equivalent of various aromatic isomers in dimethyl sulfoxide (DMSO) solutions of phenazine derivatives (DMSO, [c] = 2.0×10 -5 M) fluorescence signal and found that ( Figure 1 ), when in a solution of compound DCP (2.0×10 -5 When p-PD was added to the DCP solution (M), the fluorescence emission intensity increased. Furthermore, under 365nm UV illumination, the fluorescence color of DCP changed from light blue to dark blue, which was clearly visible to the naked eye. The addition of other structures had no significant effect on the fluorescence spectrum of the DCP solution. These experiments demonstrate that DCP can selectively identify p-PD with high sensitivity.
[0009] 2. Fluorescence titration experiment of phenazine derivative DCP and p-PD Pipette 2.0 mL of DMSO solution of DCP (2.0 × 10-5 mol·L -1 ) in a quartz cell, and gradually add p-PD DMSO solution by cumulative addition method, and measure its fluorescence emission spectrum at 25℃ ( Figure 2 Titration experiments showed that the fluorescence intensity of DCP was affected by the concentration of p-PD. It increased and blue-shifted with the increase of p-PD concentration. At the same time, there was a good linear relationship between the fluorescence intensity of DCP at 450 nm and the p-PD concentration (R 2 = 0.988), and the detection limits (LODs) of DCP for p-PD calculated according to the 3σ / S method were 3.96×10 -8 This indicates that DCP can selectively identify p-PD with fluorescence and has high sensitivity. Therefore, DCP has potential application value in p-PD detection.
[0010] Figure 3 The fitted straight line plot shows the fluorescence intensity of DCP in the presence of different p-PD concentrations (0 to 2.75 times the molar concentration of DCP). The fitted straight line plot clearly reflects the changing trend of DCP fluorescence intensity within the p-PD concentration range of 0 to 2.75 times. The fitted straight line plot shows a good linear relationship between the fluorescence intensity of DCP and p-PD at 0 to 2.75 times the p-PD equivalent, as shown below: Y=-233.14X+319.46;R 2 =0.988; Where, Y is the fluorescence intensity of DCP solution (au); X——p-PD molar concentration (mol / L) / DCP molar concentration (mol / L).
[0011] 3. Anti-interference performance test of aromatic isomers of phenazine derivatives DCP and p-PD In order to determine the detection effect of DCP on p-PD, we conducted the following test: 2 mL of DCP-containing dimethyl sulfoxide solution was added to two sets of 10 mL colorimetric tubes, and then 2 µL of dimethyl sulfoxide solution of various aromatic isomers (0.1 mol·L −1 ). 2µL p-PD was added to the other group, and dimethyl sulfoxide solutions (0.1mol·L) of o-phenylenediamine, m-phenylenediamine, p-phenylenediamine, catechol, resorcinol, hydroquinone, phthalic acid, isophthalic acid, terephthalic acid, o-chloroaniline, m-chloroaniline, and p-chloroaniline were added to each colorimetric tube. -1 ) 2µL. Mix the above solutions evenly and observe.
[0012] After the solution was allowed to stand, its fluorescence emission spectrum was measured at 25°C. When other substances were added to the DCP solution simultaneously with p-PD, its fluorescence intensity was barely affected. Compared to the experimental results with only p-PD added, the absorbance of other competing analytes was almost identical. Therefore, based on the experimental data, it can be concluded that DCP as a probe has extremely high selectivity for p-PD and excellent resistance to other potentially interfering substances (see Figure 1 and Figure 4 ).
[0013] 3. Preparation of test strips and detection of p-PD In order to study the application performance of DCP on p-PD, we conducted the following experiment: First, the filter paper was soaked in a DCP concentration of 2×10 -5 The DCP was then immersed in a 50% DMSO solution for 4 hours to allow DCP to fully penetrate the filter paper. The soaked filter paper was then air-dried to produce a simple test paper. The DCP-loaded test paper was irradiated with a 365 nm UV lamp. Under irradiation, the test paper showed a light blue fluorescence. When the p-PD solution was added, the blue fluorescence color of the test paper was significantly deepened ( Figure 5 This color change also deepens with increasing p-PD concentration. This demonstrates that the DCP-based test paper has sensitive detection capabilities and can easily detect the presence of p-PD.
[0014] Compared with the prior art, the present invention has the following beneficial effects: 1. The phenazine derivatives of the present invention realize the fluorescence detection of p-PD. The phenazine derivatives can highly selectively identify p-PD by fluorescence, showing good single selectivity and anti-interference ability, low detection limit (0.396 nM) and high sensitivity.
[0015] 2. The present invention prepares a portable test strip based on phenazine derivatives, which can realize real-time detection of p-PD. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] Figure 1 This is the fluorescence spectrum of the phenazine derivative DCP in dimethyl sulfoxide solution after adding p-PD; Figure 2 This is the fluorescence titration diagram of the phenazine derivative DCP in dimethyl sulfoxide solution; Figure 3 is the fluorescence linear fitting diagram of the phenazine derivative DCP; Figure 4 p-PD was added to the dimethyl sulfoxide solution of phenazine derivative DCP, and different aromatic isomers were added to the solution to produce fluorescence anti-interference images; Figure 5To test the application effect of the paper strip for p-PD detection; Figure 6 is the hydrogen spectrum of intermediate 1; Figure 7 is the mass spectrum of intermediate 1; Figure 8 is the hydrogen spectrum of intermediate 2; Figure 9 is the mass spectrum of intermediate 2; Figure 10 is the hydrogen spectrum of the phenazine derivative DCP; Figure 11 This is the mass spectrum of the phenazine derivative DCP. Specific implementation methods Example 1 Preparation of Phenazine Derivative Sensing Molecule DCP (1) Synthesis of intermediate compound 1: o-phenylenediamine (0.462 mmol, 0.05 g) and 2,5-dihydroxy-1,4-benzoquinone (0.506 mmol, 0.071 g) were added to 20 mL of water, refluxed at 100 °C for 24 h, filtered under reduced pressure, and the solid was dissolved in a hot alkaline solution. The solution was filtered while hot, and the pH of the solution was adjusted to 3-4. The solution was cooled and allowed to stand, and filtered under reduced pressure to obtain intermediate compound 1.
[0017] (2) Synthesis of intermediate compound 2: Potassium carbonate (21.706 mmol, 3 g) and benzyltriethylammonium chloride (0.439 mmol, 0.1 g) were used as co-catalysts, intermediate 1 (7 mmol, 1.5 g) and ethyl bromoacetate (16 mmol, 2.5 g) were added to 50 mL of acetonitrile and refluxed at 85°C for 15 h under nitrogen protection. The mixture was filtered while hot to remove inorganic salts, extracted with dichloromethane and distilled water, dried over anhydrous sodium sulfate, and the organic phase was separated by column chromatography to obtain a light yellow solid, which was intermediate compound 2.
[0018] (3) Synthesis of compound DCP: The intermediate compound 2 (2.6 mmol, 1 g) and sodium hydroxide (15.6 mmol, 0.624 g) were added to 20 mL DMF and reacted at 120 °C for 20 h. The mixture was acidified to 2-3 with hydrochloric acid, filtered under reduced pressure, and recrystallized from ethanol to obtain the final compound DCP. The hydrogen spectrum and mass spectrum of the intermediate and DCP are shown in Figure 2. Figure 6-11 shown.
[0019] Example 2 Fluorescence detection of p-PD using the sensing molecule DCP At a concentration of 2.0×10 -5To a 0.1 M DMSO solution of the compound DCP, add 10 equivalents of o-phenylenediamine, m-phenylenediamine, p-phenylenediamine, catechol, resorcinol, hydroquinone, phthalic acid, isophthalic acid, terephthalic acid, o-chloroaniline, m-chloroaniline, and p-chloroaniline, respectively. If the fluorescence emission intensity of the DCP solution increases and its fluorescence color changes from light blue to dark blue under 365 nm ultraviolet light, it indicates that p-phenylenediamine has been added. If there is no obvious change in the fluorescence of the DCP solution, it indicates that other aromatic isomers have been added.
[0020] Example 3 Preparation of a simple test strip and detection of p-PD First, the filter paper was soaked in DCP with a concentration of 2×10 -5 The filter paper was immersed in a 300-μm dimethyl sulfoxide solution for 4 hours to allow DCP to fully penetrate the filter paper. The soaked filter paper was then air-dried to create a simple test strip. Next, the DCP-loaded test strips were illuminated with a 365 nm UV lamp. Under illumination, the test strips exhibited a light blue fluorescence, which significantly deepened when the p-PD solution was added. This color change also deepened with increasing p-PD concentration.
Claims
1. Application of phenazine derivatives in detecting p-phenylenediamine, characterized in that: The structural formula of phenazine derivatives is: 。 2. The use of the phenazine derivative according to claim 1 in detecting p-phenylenediamine, characterized in that: To the dimethyl sulfoxide solution of phenazine derivatives, dimethyl sulfoxide solutions of o-phenylenediamine, m-phenylenediamine, p-phenylenediamine, catechol, resorcinol, hydroquinone, phthalic acid, isophthalic acid, terephthalic acid, o-chloroaniline, m-chloroaniline, and p-chloroaniline were added respectively. Only the addition of p-phenylenediamine could enhance the fluorescence of the phenazine derivative solution, and the fluorescence color changed from light blue to dark blue. The addition of other aromatic isomers had no effect on the fluorescence of the phenazine derivative solution, realizing the single selective fluorescence detection of p-phenylenediamine.
3. The use of the phenazine derivative according to claim 1 in detecting p-phenylenediamine, characterized in that: When the molar concentration of the p-phenylenediamine solution is in the range of 0 to 0.75 times the molar concentration of the phenazine derivative solution, the linear relationship between the fluorescence intensity of the phenazine derivative solution and the concentration of p-phenylenediamine is as follows: Y=-233.14X+319.46;R 2 =0.988; Wherein, Y is the fluorescence intensity of the phenazine derivative solution; X——molar concentration of p-phenylenediamine solution / molar concentration of phenazine derivative solution; The fluorescence intensity of the phenazine derivative solution was measured and substituted into the linear relationship to realize the concentration detection of the p-phenylenediamine solution.
4. The use of phenazine derivatives in the preparation of phenazine derivative fluorescence detection test strips, characterized in that: The structural formula of phenazine derivatives is: 。