Method for preparing a ratiometric fluorescent sensor for phycoerythrin based on a magnetic molecularly imprinted core-shell polymer

10895,536

16/620690

March 14, 2019

A method for preparing a ratiometric fluorescent sensor for phycoerythrin based on a magnetic molecularly imprinted core-shell polymer is provided. With Fe3O4 magnetic nanoparticles as the core, blue fluorescence-emitting carbon quantum dots (B-CDs) are coupled on the surfaces of Fe3O4 magnetic nanoparticles, and SiO2 shells carrying template molecules (phycoerythrin) are grown on the surfaces of Fe3O4/B-CDs. Then, the molecularly imprinted polymer SiO2-MIPs are obtained by eluting the template molecules, that is, Fe3O4/B-CDs/SiO2-MIPs are obtained. Fluorescence emission spectra of the dispersion of Fe3O4/B-CDs/SiO2-MIPs in the presence of different concentrations of phycoerythrin are measured. By fitting the linear relationship between the ratios Iphycoerythrin/IB-CDs of fluorescence emission peak intensities of phycoerythrin and B-CDs and the molar concentrations of phycoerythrin, the ratiometric fluorescent sensor for phycoerythrin is constructed.

Qingdao University (Qingdao, CN)

QINGDAO UNIVERSITY (Qingdao, CN)

1. A method for preparing a ratiometric fluorescent sensor for phycoerythrin based on a magnetic molecularly imprinted core-shell polymer, comprising the following steps: (1) preparation of aminated blue fluorescence-emitting carbon quantum dots (B-CDs) comprising: mixing 0.3 mL of 1,4-dioxane with 25 mL of catechol solution by ultrasound to obtain a first solution; transferring the first solution to a 50 mL high-pressure reactor with a polytetrafluoroethylene liner; heating the first solution in the high-pressure reactor for a first reaction at 180° C. for 12 hours to obtain a dark brown mixture; diluting the dark brown mixture with 20 mL of double-distilled water and centrifuging at 12,000 rpm to remove larger particles to obtain a supernatant; filtering the supernatant through a 0.4 m micro-filtration membrane to obtain a filtrate, and dialyzing the filtrate through a dialysis bag with a molecular weight cut-off of 1000 Da to remove unreacted experimental materials to obtain a second solution in the dialysis bag; collecting the second solution and subjecting to a rotary evaporation to remove 90% of water; and drying in a vacuum to obtain the B-CDs; and then dispersing the dried B-CDs in water to obtain a B-CDs aqueous dispersion; (2) preparation of carboxylated Fe 3 O 4 magnetic nanoparticles comprising: adding ferric chloride and ferrous chloride with a molar ratio of 2:1 into a 250 mL reaction flask to prepare a 100 mL mixed solution; adding 10 mL of ammonia water with a mass concentration of 25% to the mixed solution into the 250 mL reaction flask under N 2 protection to obtain a third solution; stirring the third solution rapidly to cause a second reaction; adjusting pH of the third solution to alkaline with HCl after 10 minutes of the second reaction; adding 10 mL of trisodium citrate solution to the third solution to obtain a fourth solution; and then placing the 250 mL reaction flask containing the fourth solution in a water bath at 80° C. and continuously stirring the fourth solution for a third reaction for 30 minutes to obtain a first reaction production; obtaining the carboxylated Fe 3 O 4 magnetic nanoparticles by centrifugation, washing and drying of the first reaction product and dispersing the carboxylated Fe 3 O 4 magnetic nanoparticles in water to prepare a Fe 3 O 4 aqueous dispersion; (3) preparation of magnetic molecularly imprinted core-shell polymers comprising: adding 2 mL of the B-CDs aqueous dispersion to 18 mL of aqueous dispersion containing 0.8 mL of the Fe 3 O 4 aqueous dispersion to obtain a fifth solution; stirring the fifth solution for a period of 30 minutes, and then adding phycoerythrin and 20 μL of 3-aminopropyltriethoxysilane to the fifth solution to obtain a sixth solution and allowing reaction of the sixth solution to proceed for 1 hour; then adding 40 μL of ammonia water and 40 μL of tetraethyl silicate to the sixth solution to obtain a seventh solution, and allowing reaction to proceed in the seventh solution away from light for 12 hours to obtain a second reaction product; centrifuging the second reaction product; and washing three times with a solution consisting of ethanol and acetonitrile at a volume ratio of 8:2 to remove the template molecules to obtain a product, and obtaining Fe 3 O 4 /B-CDs/SiO 2 magnetic molecularly imprinted core-shell polymer (Fe 3 O 4 /B-CDs/SiO 2 -MIPs) from the product by centrifugation, washing and drying; and then dispersing the Fe 3 O 4 /B-CDs/SO 2 -MIPs in water to provide a Fe 3 O 4 /B-CDs/SiO 2 -MIPs aqueous dispersion; and (4) adding a plurality of molar concentrations of phycoerythrin to the Fe 3 O 4 /B-CDs/SiO 2 -MIPs aqueous dispersion to form a plurality of homogeneous mixtures; incubating the plurality of homogeneous mixtures away from light for 5 minutes and measuring fluorescence emission spectra of the plurality of homogeneous mixtures; and by fitting a linear relationship between ratios I phycoerythrin /I B-CDs of fluorescence emission peak intensities of the phycoerythrin and the B-CDs and the plurality of molar concentrations of the phycoerythrin, the ratiometric fluorescent sensor for the phycoerythrin is constructed; wherein, in step (3), the mass concentration of the B-CDs is 1-10 mg/mL, the mass concentration of the carboxylated Fe 3 O 4 magnetic nanoparticles is 5-20 mg/mL, and the plurality of molar concentrations of the phycoerythrin in step (4) is 0.5-1 M.

2. The method for preparing the ratiometric fluorescent sensor for the phycoerythrin based on the magnetic molecularly imprinted core-shell polymer according to claim 1 , wherein, in step (1), a size of each of the aminated B-CDs is 1-5 nm.

3. The method for preparing the ratiometric fluorescent sensor for the phycoerythrin based on the magnetic molecularly imprinted core-shell polymer according to claim 1 , wherein, in step (2), a size of each of the carboxylated Fe 3 O 4 magnetic nanoparticles is 10-30 nm.

4. The method for preparing the ratiometric fluorescent sensor for the phycoerythrin based on the magnetic molecularly imprinted core-shell polymer according to claim 1 , wherein, in step (4), a linear detection range of the molar concentrations of the phycoerythrin is 1-500 nM, and a detection limit of the phycoerythrin is 1-10 nmol/L.

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