Fluorescent probe for detecting hypochlorous acid and preparation method thereof
A technology for detecting hypochlorous acid and fluorescent probes, applied in the field of fluorescent probes, can solve problems such as poor water solubility and inability to truly apply biological systems, and achieve the effects of rapid action, high sensitivity and strong specificity
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Embodiment 1
[0031] Example 1: Preparation method of fluorescent probe for detecting hypochlorous acid
[0032] In a 25mL round bottom flask, add dimethylaminothioformyl chloride (1.26g, 10.0mmol), dissolve in 6.0mL dry DMF, and add 7-hydroxyphenoxazinon sodium salt (0.6g, 2.5mmol). ) And potassium carbonate (0.52g, 3.8mmol), stirred at room temperature under nitrogen atmosphere for 30h. The resulting brown-yellow slurry was diluted with 350-400 mL saturated sodium chloride solution, and a red precipitate was salted out. After vacuum drying, it is separated by silica gel column (CH 2 Cl 2 Used as eluent) to obtain 0.375 g of red powder (target product, fluorescent probe for detecting hypochlorous acid). (Yield 50%).
[0033] ESI-MS: C 15 H 12 N 2 O 3 S, m / z: 301.1 (M+1) + .
[0034] 1 H NMR(CDCl 3 ): δ3.388(s, 3H, NCH 3 ), 3.477(s, 3H, NCH 3 ), 6.331 (d, J = 1.2 Hz, 1H, ArH), 6.866 (q, J = 6.0 Hz, 1H, ArH), 7.093 (q, J = 3.4 Hz, 1H, ArH), 7.120 (d, J = 2.4 Hz, 1H, ArH), 7.442 (d, J=10.0 Hz, 1...
Embodiment 2
[0036] Example 2: Description of the chemical properties of the fluorescent probe for detecting hypochlorous acid and its application in microscopic fluorescence imaging of endogenous HClO in cells
[0037] 1. Probe in neutral aqueous solution 1 The detection performance test:
[0038] (1) Preparation steps of test solution:
[0039] In a 10mL graduated test tube, add 5.0mL of 0.2mol / L pH 7.40 phosphate buffer, a certain amount of standard NaClO solution (previously calibrated accurately by spectrophotometry, see literature: M.Mazda and DWMargerum.Inorg .Chem. 1994, 33, 118.), diluted to the mark with three times distilled water. Shake well. Add 0.1mL to a concentration of 1.0×10 -3 mol / L methanol stock solution of fluorescent probe for detecting hypochlorous acid, shake well. Perform relevant measurements immediately. Operate as above, but do not add hypochlorous acid or other active oxygen detection hypochlorous acid fluorescent probe solution, it is the preparation of blank te...
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