Preparation method and application of imprinted magnetic nanoparticle PQQ-DA
A magnetic nanoparticle, PQQ-DA technology, applied in the field of biological analysis, can solve problems such as difficult detection and insufficient specific selectivity
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Embodiment 1
[0054] Example 1 Preparation of PQQ-DA imprinted magnetic nanoparticles, the steps are:
[0055] (1) Fe 3 o 4 Preparation of magnetic nanoparticles:
[0056] 100mL 1.25mM FeSO 4 ·7H 2 O was mechanically stirred at a speed of 800 rpm under the protection of nitrogen, and the pH was adjusted to 10.0 with 6M NaOH solution. After 1 hour, the magnetic precipitate was separated, and the black Fe was adsorbed by a magnet. 3 o 4 Nanoparticles, pour off the upper liquid, wash three times with double distilled water and ethanol, and then disperse and suspend in ethanol solution to obtain 5.0 g / L nano-Fe 3 o 4 solution.
[0057] (2) Magnetic Fe 3 o 4 Coated SiO 2 preparation of
[0058] 50 mL of prepared Fe 3 o 4 Add 5 mL of ammonia water, 5 mL of double distilled water, and 10 mL of 1 mM tetramethoxysilane into the suspension, mechanically stir at 800 rpm at 60°C for 10 h; add 200 μL of 3-(methacryloyloxy)propyltrimethoxysilane to disperse In 50 mL of 10% acetic acid aque...
Embodiment 2
[0072] Example 2 Determination of PQQ-DA in tissue samples
[0073] (1) Standard curve preparation
[0074] Weigh PQQ-DA and prepare 0.02, 0.04, 0.06, 0.08, 0.1 mg / mL solutions with double distilled water, respectively take 20 mg imprinted magnetic nanoparticles and disperse them in 250 mL of the above standard solution. After shaking and adsorbing at 15°C for 1 h, a magnet was placed at the bottom of the beaker to absorb the imprinted magnetic nanoparticles, and the supernatant was discarded. After the imprinted magnetic nanoparticles were washed with deionized water, they were redispersed in 5 mL of ethanol-formic acid (V / V 6 / 1) mixed solution. After the dispersion was sonicated for 10 min, the particles were attracted to the bottom of the beaker with a magnet, and the supernatant was diluted for UPLC-MS analysis. Using the integral area to plot the concentration, the regression equation is y=1205519606X+259487 (R 2 =0.9966), X is the concentration C (mg / mL), and y is the...
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