Method for quickly detecting dibutyl phthalate in food
A fast technology of dibutyl phthalate, applied in measurement devices, instruments, fluorescence/phosphorescence, etc., can solve the problems of difficult to adapt to the rapid diagnosis of food industry, cumbersome sample processing, high monitoring costs, etc., to save detection time , the method is simple, the effect of short time-consuming
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Embodiment 1
[0053] Establishment of the inhibition curve of dibutyl phthalate
[0054] (1) Coating steps:
[0055] The coating antigen DBAP-OVA at a concentration of 5μg / mL is coated with 96-well microtiter plate, 100μL per well, placed in the incubator and incubated at 37°C for 2h, then taken out of the incubator, shaken off the solution in the well, and then washed Wash with PBST and spin dry, wash 3-5min each time, wash 3 times in total.
[0056] (2) Closing steps:
[0057] Add 1% OVA solution, 150μL per well, to block the excess part that is not coated with antigen, to avoid non-specific adsorption of antibodies on the microtiter plate. After incubating at 37°C for 30 minutes, take it out of the incubator, shake off the solution in the well, and then wash as above.
[0058] (3) Competition steps:
[0059] Add 50μL of DBP standard solution and 50μL of fluorescent substance-labeled DBP antibody to each well to cause a competitive reaction. After incubating for 2.5 hours at 37°C, take it out of ...
Embodiment 2
[0068] Detecting the content of dibutyl phthalate in whole milk and adding standard recovery experiment
[0069] The pretreatment steps of whole milk: Weigh 2.0 g of whole milk solid powder (purchased from Wuhu supermarket), dissolve it in 10 mL of hot (70-90°C) distilled water, and wait for it to cool. Add 2 mL of absolute ethanol to the above 10 mL solution, shake for 5 min, and then centrifuge at 10,000 rpm at low temperature (0-4° C.) for 10 min. After centrifugation, take out the test tube, remove the upper fat layer, transfer the middle solution layer to a clean glass container, and adjust the pH of the solution to 7.0 with 1mol / L NaOH or 1mol / L HCl. Store at low temperature (0-4℃) )under.
[0070] 1. Detect the content of dibutyl phthalate in whole milk
[0071] except
[0072] (3) Competition steps:
[0073] Add 50μL of treated whole milk solution and 50μL of fluorescent substance-labeled DBP antibody solution to each well to cause a competitive reaction. After incubating fo...
Embodiment 3
[0081] Example 3: Detecting the content of dibutyl phthalate in liquid milk tea and adding standard recovery experiment
[0082] Pretreatment steps of liquid milk tea:
[0083] Add 2 mL of absolute ethanol to 10 mL of purchased liquid milk tea (purchased in a Wuhu supermarket) solution, shake for 5 min, and then centrifuge at 10,000 rpm at low temperature (0-4° C.) for 10 min. After centrifugation, take out the test tube, remove the upper fat layer, transfer the middle solution layer to a clean glass container, and adjust the pH of the solution to 7.0 with 1mol / L NaOH or 1mol / L HCl. Store at low temperature (0-4℃) )under.
[0084] 1. Detect the content of dibutyl phthalate in milk tea
[0085] (1) Coating steps:
[0086] The coating antigen DBAP-OVA at a concentration of 5μg / mL is coated with 96-well microtiter plate, 100μL per well, placed in the incubator and incubated at 37°C for 2h, then taken out of the incubator, shaken off the solution in the well, and then washed Wash with PB...
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