A kind of bifunctional fluorescent probe for detecting viscosity and pH and its preparation and application
A fluorescent probe, dual-function technology, applied in the field of fluorescent probes, can solve the problem of scarcity of dual-function probes
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Embodiment 1
[0050] Embodiment 1: the preparation of probe (I)
[0051]
[0052] Compound II was purchased from Anaiji Company.
[0053] Compound III is a disclosed compound, which is directly referred to in this example (Wu Y Y, Yu W T, Hou T C, et al. A selective and sensitive fluorescent albumin probe for the determination of urinary albumin [J]. Chemical Communications, 2014, 50( 78): 11507-11480.) in the method preparation.
[0054] Under inert gas (nitrogen) protection, compound II, compound III and sodium methylate are added in acetonitrile solvent, the ratio of the amount of compound II, compound III and sodium methylate substance is 1:1.2:0.2, wherein compound II is 1mmol, The amount of acetonitrile was 10 mL, the reaction was stirred at 35°C for 12 h, the reactant was concentrated under reduced pressure, and purified by silica column chromatography using ethyl acetate / petroleum ether (v / v, 1:5) to obtain the probe (I) (yield 60%), its proton nuclear magnetic spectrum is refe...
Embodiment 2
[0056] Example 2: UV-Vis absorption spectrum test of probe (I) (5 μM) in PBS and 90% glycerol by volume fraction.
[0057] Accurately weigh a certain amount of probe (I) (prepared in Example 1), prepare a probe mother solution with a concentration of 1 mM with dimethyl sulfoxide, pipette 2 μL and add it to 0.398 mL glycerol / PBS buffer solution (glycerol / PBS, v:v, 1:9), and then added to the 96-well plate, then measure the UV-visible light absorption spectrum of the probe (I).
[0058] Fluorescence spectrum see image 3 . The experimental results show that when the viscosity of the buffer solution increases, the absorption of the probe (I) at 480nm is continuously enhanced, indicating that the probe (I) has a recognition effect on the viscosity.
Embodiment 3
[0059] Example 3: Probe (I) under the condition of DMSO / PBS buffer (v / v=1 / 199), pH=7.4, the fluorescence performance test of the change of the fluorescence intensity of the probe with the viscosity.
[0060] Accurately weigh a certain amount of probe (I) (prepared in Example 1), prepare a probe mother solution with a concentration of 1 mM with dimethyl sulfoxide, and pipette 2 μL of PBS buffer solution added to 0.398 mL of different viscosity values (the value of final viscosity is respectively 100,200,300,400,500,600,700,800,950), was added in 96 orifice plate at 37 ℃, then measure the change of the fluorescent intensity of probe (I), and Make related linear curves.
[0061] Fluorescence spectrum see Figure 4 , 5 . The data show that as the viscosity of the buffer increases, the fluorescence intensity at the excitation wavelength of 480nm and the emission wavelength of 535nm increases by nearly 100 times. At the same time, ln (fluorescence intensity) is the ordinate, and...
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