A method for enrichment, purification and detection of afb 1 Modified inverse opal photonic crystal microspheres and its preparation method and application
A technology of photonic crystal microspheres and inverse opal, which is applied in measurement devices, instruments, biological material analysis, etc., can solve the problems of high price and complicated process of immunoaffinity column, and achieve low price of raw materials, good effect and accurate detection. Effect
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Embodiment 1
[0044](1) Preparation of inverse opal photonic crystal microspheres: the syringe containing 18% mg / mL silica (20nm) and 9.4% mg / mL polystyrene (280nm) mixed nanoemulsion was used as the water phase, and the The syringe of pure methyl silicone oil is used as the oil phase, the syringe is connected to the micro-flow injection pump, the needle of the syringe is connected to the three-way pipe, and the flow rate is controlled (the flow rate of silicon dioxide and polystyrene emulsion is 14ml / h, and the oil phase is 12ml / h), so that the oil phase Truncating the aqueous phase forms water-in-oil microspheres of suitable size.
[0045] Put the truncated water-in-oil microspheres in a receiver filled with methyl silicone oil, and heat them at 60°C until the water evaporates completely, the nanoparticles inside the microspheres self-assemble, and the microspheres become solid and dry Curing is complete. Recover the excess methyl silicone oil, wash the residual methyl silicone oil and s...
Embodiment 2
[0049] Standard AFB 1 Enrichment purification detection:
[0050] Put the modified inverse opal photonic crystal microspheres prepared in Example 1 in a centrifuge tube, 30 μL per tube, take 5 tubes and add the AFB 1 The standard solution was used for antigen-antibody binding reaction and shaken at 37°C for 1 hour. After binding for 1 h, wash with PBS solution (pH 7.4) three times. Afterwards, 800 μL of methanol was used for four times for 10 minutes to shake at room temperature for elution, and the eluents were collected and placed in 5 mL glass reagent bottles, and placed in a 50°C water bath to blow dry with nitrogen flow. Add 300 μL of trifluoroacetic acid derivatization reagent for derivatization, and derivatize at room temperature for 3 min. After derivatization, place in a 50°C water bath and blow dry with nitrogen flow. After drying, add 200 μL of methanol solution (filtered through a 220nm filter head) to dissolve, and then use HPLC to detect. : methanol: water ...
Embodiment 3
[0053] Processing of grain samples:
[0054] Select rice, wheat, corn three agricultural product samples for AFB 1 The spike recovery experiment. Purchase three kinds of samples from the market, use a pulverizer to grind, and pass through a 20-mesh sieve. Accurately weigh 5.0g samples, weigh three parts of each sample, place them in a 125mL conical flask with a stopper, and prepare 1000ng / mL AFB 1 Methanol solution, add 0.25mL, 1.25mL, 2.5mLAFB to the sample respectively 1 methanol solution. Shake the sample with the AFB 1 Mix the toxins and place in a ventilated place to evaporate the methanol to dryness. Add 1 g of sodium chloride and 25 mL of sample extract (methanol: water = 80:20) to each of the volatilized samples, mix well, place in a homogenizer for homogenization for 1 min, and then shake and extract at room temperature for 30 min. Filtrate with Whatman4 qualitative filter paper, accurately take 10mL of the filtrate to pass through a 450nm filter head, collect t...
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