Preparation method of golden flower nanoparticle immune chromatography test strip capable of simultaneously detecting deoxynivalenol and fumonisins B1 in grains
A technology of deoxynivalenol and nanoparticles, applied in measuring devices, analytical materials, instruments, etc., to achieve broad market prospects, simple operation, and high sensitivity
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Embodiment 1
[0037] Example 1. Simultaneous fast detection of DON and FB 1 Preparation of trial strips.
[0038] 1. DON and FB 1 Preparation of monoclonal antibody-colloidal gold labels.
[0039] (1) Preparation of golden flower nanoparticles.
[0040] Put 75 μL of chloroauric acid solution with a concentration of 100 mM (1g of chloroauric acid in 25ml of deionized water) into a conical flask containing 30 mL of deionized water, stir vigorously and heat to boiling, add 900 μL of a mass fraction of 1 % sodium citrate aqueous solution and keep boiling until the color of the solution turns red, and heat for 10 min. Bring to room temperature for later use.
[0041] For a typical synthetic sea urchin-like nanoparticle, 25 μL of aqueous chloroauric acid (100 mM) was placed in 9.6 mL of deionized water with vigorous stirring, followed by the addition of 50 μL of gold seeds, 22 μL of 1% citric acid Sodium, and 1000 μL of 30 mM hydroquinone (33 mg added to 10 mL of deionized water), the solut...
Embodiment 2
[0050] Example 2. DON and FB in the sample 1 simultaneous detection.
[0051] DON and FB in rice, wheat and corn 1 simultaneous detection.
[0052] (1) Sample pretreatment and testing.
[0053] Weigh 1.0 g of pulverized samples to be tested (such as rice, wheat and corn) into a conical flask, add 100 mL of distilled aqueous solution, shake well for 3-5 min, filter with qualitative filter paper, and collect the filtrate; then take 0.5 mL of Add the filtrate to 1 mL of distilled water, mix well, and use it as the solution to be tested.
[0054] Take the above solution to be tested, add 2 drops of anionic surfactant, 0.5% Tween-20 in PBS (0.05M) solution, as the sample solution.
[0055] Take the above-mentioned sample liquid and drop it into the sample hole of the test strip, and the test result can be observed after 5 minutes.
[0056] (2) Result analysis.
[0057] When the test strip shows three red lines, it means: the content of DON is lower than 1000 μg / kg, and the c...
Embodiment 3
[0058] Example 3. (Application example).
[0059] 1. Specificity experiment.
[0060] According to the method described in Example 2, the mycotoxins such as 3-AC-DON, Nivalenol, T-2, HT-2, AFB, OTA, Citrinin, α-Zearalenol and β-Zearalenol were tested with this patent test strip There were three red lines in the test, and the result was that the test strip had no crossability with 3-AC-DON, Nivalenol, T-2, HT-2, AFB, OTA, Citrinin, α-Zearalenol and β-Zearalenol and other mycotoxins reaction.
[0061] 2. Sensitivity experiment.
[0062] Using the method described in Implementation 2, use test strips to detect DON and FB at concentrations of 0, 0; 2, 2; 5, 5; 10, 10; and 20, 20 ng / mL, respectively 1 Mixed standards, repeated 10 times, wherein when the DON concentration is equal to or higher than 5 ng / mL, the DON detection line does not show red; and when the FB 1 At concentrations equal to or higher than 5 ng / mL, FB 1 The detection line does not show red. Therefore, the DO...
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