Pyrenes fluorescent probe and preparation method and application thereof
A fluorescent probe and pyrene technology, applied in the field of fluorescent probes, can solve the problems of loss of consciousness or even death, environmental pollution, human vomiting, etc., and achieve the effects of simple preparation method, simple detection method, and good cell membrane permeability.
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Embodiment 1
[0053] The preparation of embodiment 1 pyrene fluorescent probe, the steps are as follows:
[0054] Take 0.61g (2.5mmol) of acetylpyrene and dissolve it in 60mL of absolute ethanol in a 250mL two-necked flask (it can be heated appropriately to accelerate the dissolution), and then add 0.63g (3.5mmol) of 2,6 -dichloropyridine-3-carbaldehyde solution, then add 3 mL of 10% aqueous sodium hydroxide solution, and stir with a magnetic stirrer at room temperature for 20 h. After the reaction was completed, it was filtered, spotted on a plate to confirm the purity of the product, dried naturally for 3 hours, and then dried in a vacuum oven for 2 hours to obtain 0.89 g of an orange-yellow product with a yield of 88.6%.
[0055] The fluorescent probe was characterized by NMR, and the results are as follows:
[0056] 1 H NMR (600MHz, DMSO-d 6 )δ / ppm: 8.803~8.788(d, 1H, J=9.0Hz), 8.728~8.714(d, 1H, J=8.4Hz), 8.636~8.623(d, 1H, J=7.8Hz), 8.467~8.429 (m, 3H), 8.396~8.380(d, 2H, J=9.6Hz)...
Embodiment 2
[0057] The H of embodiment 2 pyrene fluorescent probes + Titration fluorescence plot
[0058] Add 30 μL pyrene-based fluorescent probe stock solution to 2 ml distilled water for H + Fluorescence titration experiments were detected on a fluorescence spectrophotometer. With the addition of the sample to be tested, the fluorescence intensity at 460nm weakens, and the fluorescence at 420nm increases gradually. Instrument parameters: the slit widths of the excitation wavelength and emission wavelength are 5.0nm and 5.0nm respectively, and the maximum excitation wavelength of the fluorescent probe solution is: λ ex 365nm and maximum fluorescence emission wavelength: λ em 460nm.
Embodiment 3
[0059] Example 3 Pyrene fluorescent probe pair H + Responsive working curve
[0060] Prepare 0.01M, 0.1M, 1M H with distilled water + Solution, 2ml distilled water and 30 μ L pyrene fluorescent probe stock solution are added in the fluorescence cuvette, carry out titration experiment (pH variation range 7.30-1.95); Take pH as abscissa, relative fluorescence intensity I 420nm / I 460nm Draw a graph for the ordinate and perform Sigmoidal fitting to obtain pK a =3.76; Obtaining the best linear response range is pH 3.10-4.66 by linear fitting, and the regression equation is: I 420nm / I 460nm =3.39-0.64*pH, the linear coefficient is R 2 = 0.9819.
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