A fluorescent probe for detecting hypochlorite and phosgene, and its preparation method and application

Abstract

The present invention belongs to the technical field of fluorescence detection, and in particular relates to a method for detecting hypochlorite (ClO ‑ ) and phosgene (COCl2) fluorescent probes and their preparation methods and applications. The probe is 7-(diethylamino)-2-oxy-1,2-dihydroquinoline-3-formaldehyde oxime (abbreviated as DOQC). 3-Formyl-7-diethylaminoquinoline-2( 1H )-ketone, hydroxylamine hydrochloride, solvent, and acid binding agent were added to the reaction bottle, stirred, and heated until the reaction was completed. The reaction progress was monitored by thin layer chromatography, and the fluorescent probe DOQC was obtained after filtration and purification. ‑ After the reaction of COCl2 and probe DOQC, the fluorescence color and intensity of the solution changed. This change has the characteristics of high selectivity and high sensitivity. ‑ , COCl2 rapid practical detection has improved new methods and application prospects.

Description

Technical Field

[0001] The present invention belongs to the technical field of fluorescence detection, and particularly relates to a fluorescent probe and a preparation method thereof and its application in detecting hypochlorite (ClO - ) and phosgene (COCl2). Background Art

[0002] Hypochlorite (ClO - ) has a valence of +1, which is the lowest valence oxygen-containing ion of chlorine. Its oxidizing property is extremely strong among the oxygen-containing acids of chlorine. Its salts and conjugate acids (HClO) are often used as disinfectants and bleaches. - It is one of the prominent reactive oxygen species in the immune system and plays a key role in physiology. However, abnormal ClO- concentration in the body also affects normal physiological functions. Therefore, real-time and simple detection methods are particularly important.

[0003] Phosgene (also known as carbonyl chloride, COCl2), as an easily available and highly active acylating agent, is widely used in the production and processing of medicines, pesticides, plastics, dyes, and raw materials. However, COCl2 can easily damage the respiratory system of organisms, leading to deterioration of lung tissue and pulmonary edema, which can directly lead to respiratory failure or even death in severe cases. Therefore, it is very meaningful to develop a rapid and sensitive method for detecting COCl2 in its production, use, storage, and transportation.

[0004] Traditional detection of ClO - There are many methods for detecting ClO or COCl2, such as iodine titration, electrochemical method, gas chromatography, etc. Although these methods have the characteristics of high selectivity, they still have the disadvantages of complex pretreatment and cumbersome operation. Compared with traditional detection methods, fluorescence analysis has the advantages of simple pretreatment, convenient operation, rapidity and high sensitivity. - Probes based on 2- or 3-COCl2 show good sensitivity and selectivity, but these probes also have disadvantages such as long synthesis routes and unsatisfactory detection. Therefore, it is still of great significance to develop new fluorescent probes that can overcome the above shortcomings. Summary of the invention

[0005] In view of the above technical problems, the present invention provides a fluorescent probe for detecting hypochlorite and phosgene, the probe is 7-(diethylamino)-2-oxy-1,2-dihydroquinoline-3-carbaldehyde oxime (DOQC), and the specific structural formula is as follows:

[0006]

[0007] The preparation method of the fluorescent probe for detecting chlorate and phosgene comprises the following steps: adding 3-formyl-7-diethylaminoquinoline-2(1H)-one, hydroxylamine hydrochloride, a solvent and an acid binding agent into a reaction bottle, stirring, heating until the reaction is completed (monitoring the reaction progress by thin layer chromatography), filtering and purifying to obtain the fluorescent probe DOQC.

[0008] 3-Formyl-7-diethylaminoquinoline-2( 1H The molar ratio of )-ketone to hydroxylamine hydrochloride is 1:1-50, preferably 1:2.

[0009] The solvent includes any one of methanol, ethanol, acetonitrile, N,N-dimethylformamide or isopropanol, preferably N,N-dimethylformamide.

[0010] The acid binding agent includes sodium hydroxide or pyridine or triethylamine, preferably triethylamine.

[0011] The heating temperature is 40-90°C, preferably 60°C.

[0012] The fluorescent probe for detecting chlorate is used as a reagent for qualitatively identifying and quantitatively detecting chlorate ions in an ethanol aqueous solution.

[0013] Another technical solution of the present invention is to use the probe for detecting phosgene (COCl2) in acetonitrile as a reagent for qualitative identification and quantitative detection of phosgene (COCl2).

[0014] The detection of hypochlorite (ClO - ) fluorescent probe, which can qualitatively identify and quantitatively detect hypochlorite (ClO - ).

[0015] The fluorescent probe for detecting phosgene (COCl2) can qualitatively identify and quantitatively detect phosgene (COCl2) in acetonitrile.

[0016] Beneficial effects of the present invention:

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] 1. The probe molecule of the present invention is prepared by a one-step reaction of diethylaminoquinolinone aldehyde and hydroxylamine hydrochloride. The reaction has the advantages of short synthesis route, mild reaction conditions, simple post-treatment, high yield, etc.

[0019] 2. The probe molecule contains an aldehyde oxime functional group, which can react specifically and quickly with hypochlorite and phosgene to generate cyanide under mild conditions. Before and after the reaction, the structure of the probe molecule changes, causing the color and fluorescence intensity of the probe molecule solution to change significantly. It has the characteristics of rapid response, high selectivity, high sensitivity, and strong anti-interference, and can be used for hypochlorite (ClO - ) and phosgene (COCl2) for qualitative and quantitative detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The DOQC prepared in Example 1 1 H-NMR spectrum.

[0021] Figure 2 The DOQC prepared in Example 1 13 C-NMR spectrum.

[0022] Figure 3 This is the mass spectrum of DOQC prepared in Example 1.

[0023] Figure 4 The fluorescence emission spectra of DOQC prepared in Example 1 in the presence of different ions.

[0024] Figure 5 The DOQC prepared in Example 1 with / without ClO - Fluorescence intensity plots of DOQC in the presence of various other analytes.

[0025] Figure 6 The DOQC prepared in Example 1 and hypochlorite (ClO - ) concentration changes and the fluorescence emission spectra of DOQC and hypochlorite (ClO - ) part of the concentration linear relationship diagram b).

[0026] Figure 7 The DOQC prepared in Example 1 and hypochlorite (ClO - ) The relationship between fluorescence emission intensity and time during response.

[0027] Figure 8 The DOQC prepared in Example 1 was used to detect hypochlorite (ClO - ) and pH.

[0028] Fig. 9 The fluorescence emission spectra of DOQC prepared in Example 1 for phosgene (COCl2) among different analytes.

[0029] Fig.10a) is the fluorescence emission spectrum of DOQC prepared in Example 1 and the concentration change of phosgene (COCl2) and b) is the linear relationship diagram of the concentration change of DOQC and phosgene (COCl2) part.

[0030] Fig.11 This is a graph showing the relationship between the fluorescence intensity and time of the DOQC prepared in Example 1 when reacting with phosgene (COCl2). DETAILED DESCRIPTION

[0031] The present invention is further described below with reference to embodiments, but the scope of protection claimed by the present invention is not limited to the scope described in the embodiments.

[0032] Example 1

[0033] Preparation of 7-(Diethylamino)-2-oxo-1,2-dihydroquinoline-3-carbaldehyde oxime (DOQC)

[0034] 3-Formyl-7-diethylaminoquinoline-2( 1H )-ketone (2.36 g, 10.01 mmol) was added to a 50 mL two-necked bottle, followed by hydroxylamine hydrochloride (1.39 g, 20.08 mmol), and finally 8.0 mL of N,N-dimethylformamide and 2 mL of triethylamine. The mixture was stirred at 60 °C for 4 h. After the reaction was complete (TLC monitoring), the mixture was naturally cooled to room temperature, 20.0 mL of water was added, and the mixture was filtered. The obtained solid was recrystallized from N,N-dimethylformamide / water and dried to obtain 2.34 g of a yellow solid with a yield of 90%. 1 HNMR (400 MHz, DMSO) δ 11.42 (s, 1H), 8.13 (s, 1H), 8.04 (s, 1H), 7.49 (d, J =9.0 Hz, 1H), 6.64 (dd, J = 9.0, 2.3 Hz, 1H), 6.47 (d, J = 2.1 Hz, 1H), 3.40 (d, J =7.2 Hz, 4H), 1.13 (t, J = 7.0 Hz, 6H). 13 C NMR (101 MHz, DMSO) δ 161.67, 149.96, 144.18, 141.87, 134.71, 117.37, 109.97, 109.14, 94.60, 44.51, 12.92.

[0035]

[0036] Example 1-1 obtained 2.34 g of yellow solid, with a yield of 90%. mp: 151.5-152.3 °C. 1 H NMR (400MHz, DMSO) δ 11.43 (s, 1H), 11.00 (s, 1H), 8.13 (s, 1H), 8.04 (s, 1H), 7.49(d, J = 9.0 Hz, 1H), 6.64 (dd, J = 9.0, 2.3 Hz, 1H), 6.47 (d, J = 2.1 Hz, 1H), 3.40 (d, J = 7.2 Hz, 4H), 1.13 (t, J = 7.0 Hz, 6H). 13 C NMR (100 MHz, DMSO) δ 161.67, 149.96, 144.18, 141.87, 134.71, 117.37, 109.97, 109.14, 94.60, 44.51, 12.92.

[0037] Example 2

[0038] Preparation of test solution

[0039] (1) Preparation procedure of stock solution:

[0040] DOQC detection of ClO - DOQC was dissolved in DMSO to prepare a 1.0 mmol / L stock solution, and the stock solution was diluted to 10 μmol / L in ethanol / water (v / v, 3:2) for spectral testing. The following spectral tests were all performed in an ethanol / water (v / v, 3:2) system. The concentration of anions and other analytes was 10 mmol / L. Solutions of different pH values ​​of ethanol / water (v / v, 3:2) were prepared and monitored with a Lei magnetic pH meter. The fluorescence emission spectrum was measured (360 nm as the excitation wavelength). The above operation was performed without adding the sample solution to be tested, which was the preparation of the blank test solution.

[0041] DOQC detection of COCl2: DOQC was dissolved in DMSO to prepare a 1.0 mmol / L stock solution, which was diluted to 10 μmol / L in acetonitrile for spectral testing. The following spectral tests were all performed in acetonitrile. The concentration of other analytes was 10 mmol / L. Fluorescence emission spectra were measured (360 nm as the excitation wavelength). The above operation was performed without adding the sample solution to be tested, which was the preparation of the blank test solution.

[0042] (2) Detection of hypochlorite (ClO - ) of the selective experiment:

[0043] Take 23 portions of 2 mL ethanol / water (v / v, 3:2) solution and add 20 μL of the stock solution and different anions (C5H7O5CO2 - 、F - , Cl - Br - ,I - 、HCO3 - 、CO3 2- , HS - , S 2- 、SO3 2- 、HSO3 - 、S2O3 2- 、SO4 2- 、S2O8 2- 、NO2 - 、NO3 - , CN - 、SCN - 、CH3COO - 、HPO4 2- 、H2PO4 - , ClO - When the same concentration of various analytes were reacted with DOQC, the blank test solution and the DOQC solution with other analytes had a strong emission peak at 479 nm in the fluorescence spectrum, but the addition of hypochlorite (ClO - ), the fluorescence intensity of DOQC solution at 479 nm was significantly weakened. This indicates that DOQC has a strong affinity for hypochlorite (ClO - ) is highly specific and selective, see Figure 4 .

[0044] In fact, the compounds of the present application also have the above-mentioned fluorescence specific selectivity in methanol / water (v / v, 3:2) solution, isopropanol / water (v / v, 3:2) solution, butanol / water (v / v, 3:2) solution, and amyl alcohol / water (v / v, 3:2) solution. The present application only uses ethanol / water (v / v, 3:2) solution as a representative for the following experiments:

[0045] (3) Coexisting ions to hypochlorite (ClO - )Determination of the impact of:

[0046] To further investigate the effect of DOQC on hypochlorite (ClO - ) detection selectivity, and tried to detect the analyte and hypochlorite (ClO - ) coexistence on the fluorescence emission of the system, the analytes to be measured are: C5H7O5CO2 - 、F -, Cl - Br - ,I - 、HCO3 - 、CO3 2- , HS - , S 2- 、SO3 2- 、HSO3 - 、S2O3 2- 、SO4 2- 、S2O8 2- 、NO2 - 、NO3 - , CN - 、SCN - 、CH3COO - 、HPO4 2- 、H2PO4 - ,See Figure 5 . Except for ClO - Coexisting ions (HSO3 - 、S2O3 2- 、SCN - ,I - 、NO2 - , S 2- ) exists, other analyte ions will basically not interfere with ClO - Detection, but C5H7O5CO2 - 、F - To a certain extent, it will interfere with DOQC's ClO - The results show that DOQC has a significant effect on ClO - The detection has high anti-interference ability.

[0047] (4) Detection of hypochlorite (ClO - ) titration experiment:

[0048] In the fluorescence emission spectrum, linear fitting shows that hypochlorite (ClO - )Concentration curve: y=-44.28457x+4384.67714(R 2 =0.9927, hypochlorite (ClO - ) concentration range: 70~95 μM); see Figure 6 (Concentration interval is 5 μM). According to the minimum detection limit formula (LOD=3σ / b), DOQC has a significant effect on ClO in fluorescence spectrophotometry. - The detection limit is: 1.96×10 -8 mol / L.

[0049] (5) Detection of hypochlorite (ClO - ) Kinetic experiment:

[0050] The fluorescence spectra of DOQC and hypochlorite (ClO) were studied by time-dependent fluorescence spectroscopy. - ) between the reaction kinetics. Figure 7 As shown, hypochlorite (ClO) was added to an ethanol / water (v / v, 3:2) solution of DOQC. - ), the fluorescence intensity of DOQC at 479 nm was rapidly quenched, indicating a rapid response. This phenomenon indicates that DOQC and hypochlorite (ClO - ) can be used as a real-time detector for hypochlorite (ClO - ) tool. Therefore, the probe DOQC can detect hypochlorite (ClO - ).

[0051] (6) Test of fluorescence emission spectrum of probe detecting hypochlorite in response to pH change:

[0052] Take 2 mL of ethanol / water (v / v, 3:2) solution with different pH values, add 20 μL of stock solution for spectral testing, and then add 20 μL of hypochlorite (ClO - ) solution for spectral testing. In acidic and alkaline environments, DOQC has a good detection effect on the detection of hypochlorite (ClO - ) have an effect on the ion exchange rate of DOQC. - ) is more responsive, see Figure 8 .

[0053] (7) Detection of hypochlorite (ClO - ) recovery experiment:

[0054] Take two water samples, stream water and tap water, and prepare ethanol / water (v / v, 3 / 2) with ethanol. Perform DOQC on ClO - The recovery rate of spiked DOQC in ClO - Three points were randomly selected at different concentrations (70 μmol / L-100 μmol / L), and the recovery rate and relative standard deviation were calculated by fluorescence emission spectrum detection. The recovery rate of DOQC in actual water samples was 88.30%-95.81%, and the relative standard deviation was within 4%, indicating that the probe DOQC can be applied to the detection in actual water samples. See Table 1:

[0055] Table 1 DOQC for ClO in different water samples - Recovery experiment

[0056]

[0057] (8) Selective experiment for detecting phosgene (COCl2):

[0058] Take 7 portions of 2 mL acetonitrile solution, add 20 μL of stock solution and different analytes (phosphorus oxychloride, thionyl chloride, phosgene, benzoyl chloride, acetyl chloride, sulfonyl chloride) to each for fluorescence emission spectrum testing. When the same concentration of various analytes to be tested react with DOQC, in the fluorescence spectrum, the blank test solution and the DOQC solution with other analytes to be tested have a strong emission peak at 469 nm, but after adding phosgene, the fluorescence intensity of the DOQC solution at 469 nm is significantly weakened and blue-shifted to 450 nm. This shows that DOQC has a high degree of specific selectivity for the detection of phosgene, see Fig. 9 .

[0059] (9) Titration experiment for detecting phosgene (COCl2):

[0060] In the fluorescence emission spectrum, the linear fitting results in the detection phosgene (COCl2) concentration curve: y=-22.8551x +2269.9096 (R 2 =0.99816, the concentration range of phosgene (COCl2) is: 0~75 μM); see Fig.10 (Concentration interval is 5 μM). According to the minimum detection limit formula (LOD=3σ / b), DOQC has a significant effect on ClO in fluorescence spectrophotometry. - The detection limit is: 3.45×10 -8 mol / L.

[0061] (10) Kinetic experiment for detecting phosgene (COCl2):

[0062] The reaction kinetics between DOQC and phosgene (COCl2) was studied by fluorescence spectroscopy over time. Fig.11 As shown in the figure, after adding phosgene (COCl2) to the acetonitrile solution of DOQC, the fluorescence intensity of DOQC at 469 nm decreased rapidly within 30 s and reached a stable state at 60 s, indicating a rapid response. This phenomenon indicates that the rapid response between DOQC and phosgene (COCl2) can be used as a favorable detection method for real-time detection of phosgene (COCl2). Therefore, the probe DOQC can detect the presence of phosgene (COCl2) in the environment or solution in real time.

Claims

1. Fluorescent probes for the detection of hypochlorite ClO in alcohol solutions for non-disease diagnosis and treatment purposes - The structural formula of the fluorescent probe is as follows: 。 2. The use according to claim 1, characterized in that: The alcohol solution is a C1-C5 alcohol or an aqueous solution of a C1-C5 alcohol.

3. Application of fluorescent probe in the detection solvent environment of phosgene COCl2 for non-disease diagnosis and treatment purposes in an alcohol solution environment. The structural formula of the fluorescent probe is as follows: 。 4. The use according to claim 3, characterized in that: Application of fluorescent probe to detect phosgene COCl2 in acetonitrile environment or acetonitrile aqueous solution environment.

5. The use according to any one of claims 1 to 4, characterized in that: The fluorescent probe is used for qualitative identification and quantitative detection of hypochlorite ClO - Or phosgene COCl2.

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