Metal oxide nano-composite magnetic material, fabrication method, and method for linkage, enrichment, and isolation of phosphorylated species
a metal oxide and nano-composite technology, applied in the field of metal oxide nano-composite magnetic materials, can solve the problems of poor extraction capacity and low sensitivity, and achieve the effect of high selectivity and speed, and easy linkag
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first preferred embodiment
Methods for Generating Metal Oxide Nano-Composite Magnetic Materials:
[0039]magnetic iron oxide nanoparticles were prepared by the steps described as follows. Iron(III) chloride (FeCl3.6H2O, 6.48 g) was dissolved in hydrochloric acid (2 M, 12 mL) and then diluted in 100 mL of water. The mixture was degassed using a pump. Sodium sulfite (50 mL, 0.08 M) was slowly added to the solution under nitrogen with stirring at room temperature. Subsequently, ammonia (5%, 45 mL) was added slowly to the reaction solution with vigorously stirring at room temperature. The whole mixture was left to react for 30 min in a water bath at 70° C. The magnetic iron oxide nanoparticles were rinsed with deionized water three times.
[0040]The magnetic iron oxide nanoparticles in water (5 mg / mL, 40 mL) was well suspended in an air-free condition by sonication under nitrogen gas. Sodium silicate aqueous solution (0.6%, pH 9, 40 mL) was added into the solution and was vortex-mixed for 24 h at 35° C. to coat a thin...
second preferred embodiment
Methods for Linkage, Enrichment, and Isolation of Phosphorylated Species
[0043]Protein mixtures were prepared in ammonium bicarbonate solution (0.5 mL, 50 mM) containing urea (8 M), while the milk (0.25 mL) was mixed with the same solution (0.25 mL), followed by incubation at 38° C. for 30 min. A DTT (50 μL, 100 mM) prepared in ammonium bicarbonate solution (50 mM) was added into this mixture, which was maintained at 50° C. for 1 h. After cooling to room temperature, the sample solution was mixed with IAA solution (50 μL, 200 mM) prepared in ammonium bicarbonate solution (50 mM) in an ice bath. The sample vial was wrapped with aluminum foil and mixed by vortex mixing for 2.5 h at room temperature. After 2.5 h, the sample solution (sample A2) was ready for carrying out either the enrichment process by the magnetic particles or tryptic digestion.
[0044]To demonstrate the feasibility of employing this approach to a real sample, the nanoparticles of the metal oxide nano-composite magnetic...
third preferred embodiment
Methods for Fabrication of Metal Oxide Nano-Composite Magnetic Nanoparticles
[0045]FeCl3 (2 g) and FeCl2 (5.4 g) were dissolved in aqueous hydrochloric acid (2 M, 25 mL) at room temperature under sonication. After the salts had dissolved completely in the solution, the mixture was degassed using a vacuum pump and then the flask was filled with nitrogen gas. Aqueous ammonia (25%, 40 mL) was slowly injected into the mixture under nitrogen while stirring at room temperature, followed by rinsing with deionized water three times and re-suspension in ethanol (40 mL). The final concentration of the magnetic iron oxide nanoparticle suspension was 40 mg / mL.
[0046]The magnetic iron oxide nanoparticles solution (40 mg / mL, 23 mL) obtained above was sonicated with ethanol (35 mL) and deionized water (6 mL) for 10 min. Tetraethoxysilane (TEOS, 1.5 mL) was added, and the mixture was then sonicated for 10 min; aqueous ammonia (10%, 1.4 mL) was then added slowly over 5 min. The mixture was stirred in ...
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