Kit for detecting aflatoxin B1 and preparation method thereof
A kind of technology of aflatoxin and kit, which is applied in the field of kit for visual detection of aflatoxin B1 and its preparation, can solve the problems of poor acid and alkali resistance, high synthesis cost, long detection cycle, etc., and achieve small steric hindrance , Improve detection performance, strong binding effect
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Embodiment 1
[0056] The preparation method of the kit for rapid detection of aflatoxin B1 in food of the present invention comprises:
[0057] Step 1: Prepare platinum-gold silica nano-microsphere signal label and Fe3O4 nano-microsphere capture agent.
[0058] Step 2: Take 1mL of platinum silica nanosphere signal label with a particle size of 900nm and a concentration of 0.19mg / L and 1mL of a particle size of 400nm with a concentration of 1ⅹ10 -9 The mol / L iron ferric oxide nano-microsphere capture agent was mixed and placed in a glass container for 20 minutes, then ultrasonicated for 5 minutes, and then heated to 95° C. for 10 minutes to obtain the product.
[0059] Step 3: Wash the reaction product obtained in Step 2 repeatedly with a magnetic field, and disperse it into 2 mL of water.
[0060] Step 4: Take 10 μL of the product solution obtained from the reaction in step 3 and drop it into each reaction well of a 96-well reaction plate to obtain a kit for detecting aflatoxin B1 in food....
Embodiment 2
[0070] The preparation of the platinum silica nano-microsphere signal label specifically includes the following steps:
[0071] Step A1, take 1.04mL of tetrachloroauric acid with a concentration of 24mmol / L, add 7.3535mg of trisodium citrate and set the volume to 100mL, add 3mL of hydroboration with a concentration of 0.1mol / L in an ice bath under constant stirring The sodium solution was stirred for 24 hours to obtain gold nanoparticles, and the gold nanoparticles were centrifugally washed and dispersed in 100 mL of water for later use.
[0072]Step A2: Take 68mg of triethylamine and add 25mL of water, stir magnetically at 80°C for 0.5h, add 368mg of cetyltrimethylammonium bromide and 92mg of salicylic acid at 80°C for 1h, then add 4mL of orthosilicic acid After ethyl ester was reacted at 80°C for 1 hour, take it out and cool it down, then centrifuge and wash it to get the precipitate, then add 25mL hydrochloric acid methanol solution, the volume ratio of the hydrochloric aci...
Embodiment 3
[0089] The reaction plate in the kit is a 96-well reaction plate, such as figure 1 As shown, 96 holes of the same size are opened on the board body, divided into 8 rows and 12 columns, and the distance between two adjacent holes in the vertical or horizontal direction is the same. An automatically detachable box is assembled at the bottom of the reaction plate, such as figure 2 As shown, the length and width of the liner of the box are the same as that of a 96-well reaction plate, and the height is 14.35mm. Place magnets of the same size in the box.
[0090] In addition, the reaction wells of the 96-well plate store the product solution of the platinum silica microsphere signal label and the Fe3O4 nanometer microsphere capture agent.
[0091] With this design, capture, enrichment, separation and washing operations can be conveniently performed, and the time for wrapping and washing of ELISA can be reduced.
[0092] And because the magnet box in the kit can be automatically...
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