Preparation method and application of imprinted magnetic nanoparticle PQQ-DA
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A magnetic nanoparticle, PQQ-DA technology, applied in the field of biological analysis, can solve problems such as difficult detection and insufficient specific selectivity
Inactive Publication Date: 2017-10-03
JIANGSU INST OF NUCLEAR MEDICINE
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The content of PQQ-DA in the brain belongs to trace level, which is difficult to detect by general analysis methods
Conventional solid-phase extraction sorbents have insufficient specific selectivity, and it is easy to co-extract other interfering substances and target substances
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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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Abstract
The invention provides a preparation method and application of a PQQ-DA imprinted magnetic nanoparticle, belonging to the field of bioanalysis technology. According to the preparation method, Fe3O4 is used as a core and coated with SiO2; then preparation is carried out so as to obtain the PQQ-DA imprinted magnetic nanoparticle; thus, a PQQ-DA magnetic nanoparticle imprinted material with high recognizability and high selectivity on its surface. After molecular imprinting modification of a magnetic nanoparticle with specificity, the prepared imprinted magnetic nanoparticle has magnetism and specific selectivity to targets and is greatly advantaged in separation, purification and enrichment of medium-and-low-concentration compounds in brain tissue of a complex matrix. The prepared PQQ-DA imprinted magnetic nanoparticle is applied to UPLC-MS determination for analysis of trace PQQ-DA in an organism. According to the invention, UPLC-MS based on molecularly-imprinted solid-phase extraction is established in the invention, can detect the content of trace PQQ-DA in the brain of a mouse and has a detection limit of 0.2 * 10<-11> mg / mL. The PQQ-DA imprinted magnetic nanoparticle has great guidance significance to research on the occurrence and mechanism of neurological diseases.
Description
technical field [0001] The invention discloses a preparation method and application of PQQ-DA imprinted magnetic nanoparticles, belonging to the technical field of biological analysis. Background technique [0002] Dopamine (DA) is an important neurotransmitter in the brain and a signal transduction molecule that participates in many life processes and plays an important role in controlling cognitive and behavioral functions. The change of DA content in the brain is related to the occurrence of many diseases, such as autism, Parkinson's disease and schizophrenia. [0003] Pyrroloquinone quinone (PQQ) is an oxidoreductase prosthetic group, mainly distributed in prokaryotes and some plants and mammals. PQQ has an extremely important function in the nervous system. It can promote the production of nerve factors in the brain and has a neuroprotective effect on nerve cells. The rat model established with PQQ deficiency shows that the immune function and cognitive function declin...
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