Fluorescent probe for detecting sulfur dioxide derivative, detection object, and preparation method and application of fluorescent probe

Abstract

The invention relates to a fluorescent probe for detecting a sulfur dioxide derivative, a detection object, and a preparation method and application of the fluorescent probe. The sulfur dioxide derivative is a salt with sulfite or bisulfite ions, and the structural formula of the fluorescent probe is as described in the specification. In the formula, TfO<-> is trifluoromethanesulfonate ions. The fluorescent probe disclosed by the invention is a colorimetric far-infrared sulfite / bisulfite fluorescent probe, has full water solubility, can be directly dissolved in water, and does not need to be dissolved by an additional organic solvent; the response of the probe is fast, and detection can be completed within 15 s; a color development mode during detectionis a mode from a colorless state to acoloredstate, so direct naked eye detection can be realized, the detection is more visual and convenient, and the rate of false positive is low; and the lowest detection limit of sodium hydrogen sulfite is 0.11 [mu]M, and sensitivity is higher. The probe of the invention not only can meet the detection requirements of sulfite / bisulfite in organisms, but also can realize rapid detection of sulfite / bisulfite in food or medicines.

Description

technical field

[0001] The invention belongs to the field of applied biotechnology, and in particular relates to a fluorescent probe for detecting sulfur dioxide derivatives, a detection substance, a preparation method and an application. Background technique

[0002] As antibacterial agents, antioxidants and enzyme inhibitors, derivatives of sulfur dioxide (sulfite / bisulfite) are widely used in the fields of food, pharmaceutical products and beverages. However, studies have shown that excessively high concentrations of sulfite / bisulfite have greater damage to cells, tissues, and biomolecules, thereby causing many diseases, such as cancer, migraine, ischemic heart disease, and allergic reactions, etc. . Due to potential health concerns, many countries have severely restricted the amount of sulfite / bisulfite in food and beverages. For example, the Food and Drug Administration of the United States requires that the content of sulfite in food should not exceed 10mg / mL. There...

Examples

Embodiment 1

[0038] The reaction scheme of the preparation method of fluorescent probe (i.e. probe DQ) of the present invention is as follows:

[0039]

[0040] Compound 1 (212 mg, 0.5 mmol) and methyl trifluoromethanesulfonate (164 mg, 1 mmol) were dissolved in 10 mL of chloroform, the solution was stirred overnight at room temperature, and filtered with suction to obtain compound DQ (223 mg, yield 76%). Such as Figure 1-2 as shown, 1 H NMR (400MHz, DMSO-d 6 )δ(ppm): 10.15(s, 1H), 9.64(s, 1H), 9.54(d, J=6.4Hz, 1H), 9.02(d, J=8.4Hz, 1H), 8.73(d, J= 16.4Hz,1H),8.61-8.52(m,4H),8.39-8.29(m,3H),8.22-8.12(m,2H),4.75(s,3H),4.64(s,3H); 13 C NMR (100MHz, DMSO-d 6 )δ(ppm): 151.4, 150.4, 149.6, 145.3, 139.3, 138.3, 136.6, 135.9, 135.7, 131.4, 131.2, 130.4, 130.0, 129.4, 127.2, 126.5, 125.9, 120.5, 119.9, 167.4.

Embodiment 2

[0041] Embodiment 2 Probe DQ is to the absorption spectrum of bisulfite radical response and the fluorescence spectrum to sulfite radical / bisulfite radical

[0042] The probe compound DQ was dissolved in PBS solution (20 mM, pH 7.4) to form a 1 mmol / L solution. Add PBS buffer (20 mM, pH 7.4) to the pipetted probe DQ solution, and dilute to 10 μM (20 mM, pH 7.4) of the test solution. Add sulfite or bisulfite to the solution to be tested, and test the change of its fluorescence emission spectrum with a fluorescence spectrum and an ultraviolet spectrometer. The reaction mechanism is as follows: Figure 11 shown. Depend on image 3 It can be seen that the probe DQ has no fluorescence emission at 620nm, and after adding sulfite or bisulfite, the probe has a strong fluorescence emission peak at 620nm, which shows that sulfite or bisulfite can cause probe Enhancement of needle fluorescence emission. Depend on Figure 4 It can be seen that the fluorescence emission of the probe s...

Embodiment 3

[0043] Embodiment 3 Probe DQ is to the time research of bisulfite radical

[0044] The kinetics of the action of DQ on bisulfite was explored by testing the change of fluorescence intensity of probe DQ in response to bisulfite. Depend on Figure 7 It can be seen that after adding bisulfite to the probe DQ solution, the fluorescence intensity gradually increases with time, and the fluorescence intensity can reach the maximum within 15 seconds, indicating that the probe DQ can quickly detect bisulfite ions.

technical field

[0001] The invention belongs to the field of applied biotechnology, and in particular relates to a fluorescent probe for detecting sulfur dioxide derivatives, a detection substance, a preparation method and an application. Background technique

[0002] As antibacterial agents, antioxidants and enzyme inhibitors, derivatives of sulfur dioxide (sulfite / bisulfite) are widely used in the fields of food, pharmaceutical products and beverages. However, studies have shown that excessively high concentrations of sulfite / bisulfite have greater damage to cells, tissues, and biomolecules, thereby causing many diseases, such as cancer, migraine, ischemic heart disease, and allergic reactions, etc. . Due to potential health concerns, many countries have severely restricted the amount of sulfite / bisulfite in food and beverages. For example, the Food and Drug Administration of the United States requires that the content of sulfite in food should not exceed 10mg / mL. There...

Examples

Embodiment 1

[0038] The reaction scheme of the preparation method of fluorescent probe (i.e. probe DQ) of the present invention is as follows:

[0039]

[0040] Compound 1 (212 mg, 0.5 mmol) and methyl trifluoromethanesulfonate (164 mg, 1 mmol) were dissolved in 10 mL of chloroform, the solution was stirred overnight at room temperature, and filtered with suction to obtain compound DQ (223 mg, yield 76%). Such as Figure 1-2 as shown, 1 H NMR (400MHz, DMSO-d 6 )δ(ppm): 10.15(s, 1H), 9.64(s, 1H), 9.54(d, J=6.4Hz, 1H), 9.02(d, J=8.4Hz, 1H), 8.73(d, J= 16.4Hz,1H),8.61-8.52(m,4H),8.39-8.29(m,3H),8.22-8.12(m,2H),4.75(s,3H),4.64(s,3H); 13 C NMR (100MHz, DMSO-d 6 )δ(ppm): 151.4, 150.4, 149.6, 145.3, 139.3, 138.3, 136.6, 135.9, 135.7, 131.4, 131.2, 130.4, 130.0, 129.4, 127.2, 126.5, 125.9, 120.5, 119.9, 167.4.

Embodiment 2

[0041] Embodiment 2 Probe DQ is to the absorption spectrum of bisulfite radical response and the fluorescence spectrum to sulfite radical / bisulfite radical

[0042] The probe compound DQ was dissolved in PBS solution (20 mM, pH 7.4) to form a 1 mmol / L solution. Add PBS buffer (20 mM, pH 7.4) to the pipetted probe DQ solution, and dilute to 10 μM (20 mM, pH 7.4) of the test solution. Add sulfite or bisulfite to the solution to be tested, and test the change of its fluorescence emission spectrum with a fluorescence spectrum and an ultraviolet spectrometer. The reaction mechanism is as follows: Figure 11 shown. Depend on image 3 It can be seen that the probe DQ has no fluorescence emission at 620nm, and after adding sulfite or bisulfite, the probe has a strong fluorescence emission peak at 620nm, which shows that sulfite or bisulfite can cause probe Enhancement of needle fluorescence emission. Depend on Figure 4 It can be seen that the fluorescence emission of the probe s...

Embodiment 3

[0043] Embodiment 3 Probe DQ is to the time research of bisulfite radical

[0044] The kinetics of the action of DQ on bisulfite was explored by testing the change of fluorescence intensity of probe DQ in response to bisulfite. Depend on Figure 7 It can be seen that after adding bisulfite to the probe DQ solution, the fluorescence intensity gradually increases with time, and the fluorescence intensity can reach the maximum within 15 seconds, indicating that the probe DQ can quickly detect bisulfite ions.

Images