Chromium-ion ratio type fluorescence probe as well as preparation method and application thereof
A fluorescent probe, ratiometric technology, applied in the field of analytical chemistry, to achieve the effect of improving specificity and sensitivity, good water solubility, and high yield
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Embodiment 1
[0023] 1: Cr 3+ The basic operation process for the synthesis of ratiometric fluorescent probes is as follows.
[0024] 1). Synthesis of N, N'-bis(2-pyridylmethyl)piperazine
[0025] Dissolve 0.34g of anhydrous piperazine (4mmol) in 30mL of methanol, slowly add dropwise to a methanol solution of 2.0g 2-chloromethylpyridine hydrochloride (12mmol), heat up to 50°C after the dropwise addition, After reacting for 8 hours, add 0.2 g of sodium hydroxide (20 mmol) in 30 mL of methanol solution, stir for 10 min, stand for filtration, and spin dry under reduced pressure. The crude product is purified by silica gel column chromatography to obtain a yellow solid product with a yield of 62.5%. .
[0026] 2).Cr 3+ Preparation of ratiometric fluorescent probes
[0027] Dissolve 33.4mg of IR-780 iodide (0.05mmol) and 35.4mg of N,N'-bis(2-pyridylmethyl)piperazine (0.2mmol) in 15mL of DMF, under nitrogen protection, to Anhydrous DMF was used as a solvent, and the reaction was carried out ...
Embodiment 2
[0030] Embodiment 2: Solution preparation of fluorescent probe 1 interacting with chromium ions
[0031] In a 10mL volumetric flask, add 5.0mL with a concentration of 2×10 -5 mol L -1 The ethanol aqueous solution of the chromium ion ratio fluorescent probe (ethanol / water=1:4, v / v), 1.0mL1×10 -3 -1×10 -7 mol L -1 Cr 3+ Aqueous solution, dilute to volume with HEPES buffer solution. The solution to be tested thus obtained contains 1×10 -5 mol L -1 Fluorescent probes 1 and 1 x 10 -3 -1×10 -8 mol L -1 Cr 3+. The solution to be tested was stored at 4°C in the dark. A blank solution of fluorescent probe 1 was prepared under the same conditions without the addition of Cr 3+ . The water used to prepare the solutions in the experiment was double distilled water.
Embodiment 3
[0033] Fluorescent probe 1 and Cr 3+ Determination of the fluorescence spectroscopic properties of the action
[0034] Fluorescent probe 1 and Cr 3+ The fluorescence spectrum of the action, the results are as follows figure 1 . The concentration of fluorescent probe 1 was 1.0 x 10 -5 mol / L,Cr 3+ The concentrations are 0, 0.1, 0.2, 0.5, 0.8, 1, 2, 5, 8, 10 μM, the excitation wavelength is 530 nm, and the slit width is 10 nm / 10 nm. From figure 1 It can be seen that a remarkable feature of this fluorescent probe 1 is that it interacts with Cr 3+ The fluorescence emission peaks showed a large shift before and after the combination, with the Cr 3+ With the increase of the concentration, the fluorescence intensity at 561nm increases significantly, while the fluorescence intensity at 755nm decreases gradually, and the fluorescence spectrum has a blue shift. Fluorescence ratio I at 561nm and 755nm after fluorescent probe 1 interacts with chromium ions 561 / I 755 It increases...
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