A kind of test strip for detecting Yiyadan and its preparation method and application
A technology of inhibiting buds and test strips, which can be applied to measuring devices, analyze materials by chemical reaction, and analyze materials by observing the influence on chemical indicators, which can solve the problem of inability to realize rapid detection of large batches of samples. , the problem of high equipment dependence, to achieve the effect of short detection time, high sensitivity and wide application range
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Embodiment 1
[0041] Example 1 Preparation of test strips for detecting Yiyadan
[0042] The preparation method of this test strip mainly comprises the following steps:
[0043] 1) Prepare a conjugate release pad sprayed with Yiyadan monoclonal antibody-colloidal gold marker;
[0044] 2) Prepare a reaction membrane with a test line coated with Yiyadan hapten-carrier protein conjugate and a quality control line coated with goat anti-mouse anti-antibody;
[0045] 3) Assemble the conjugate release pad prepared in 1) and 2), the reaction membrane, the sample absorption pad, the water absorption pad and the PVC bottom plate into a test strip.
[0046] The following step-by-step detailed description:
[0047] 1. Synthesis of Yiyadan hapten (see attached image 3 ) and identification
[0048] Reaction a: Take 1.0 g of sulfonic maleic anhydride, add N,N-dimethylformamide (DMF) to dissolve, add 0.33 g of hydrazine sulfate, add 0.2 g of potassium hydroxide, and stir at 70°C for 4 h. Stop the react...
Embodiment 2
[0086] The detection of Yi Ya Dan in the sample of embodiment 2
[0087] 1. Sample pretreatment
[0088] Homogenize the tobacco leaf sample with a homogenizer; weigh 3.0±0.05 g of the homogenized tobacco leaf sample into a 15 mL polystyrene centrifuge tube, add 6 mL of acetonitrile, and vortex for 5 min with a vortexer; room temperature above 3000 rpm, Centrifuge for 5 min; take 2 mL of the upper organic phase into a 10 mL polystyrene centrifuge tube, and dry it in a water bath at 50-60 °C under nitrogen flow; add 100 µL of n-hexane, vortex for 30 s with a vortexer, and then add 0.5 mL Sample complex solution (0.02mol / L PBS), vortexed for 30 s to mix thoroughly; 3000 rpm at room temperature (20-25 ℃) for 5 min; remove the upper organic phase, and take 100 µL of the lower aqueous phase for analysis .
[0089] 2. Test with test strips
[0090] Use a disposable straw to absorb 2-3 drops of the sample solution to be tested and drop it vertically into the sample injection hole. ...
Embodiment 3
[0095] Embodiment 3 sample detection example
[0096] 1. Detection limit test
[0097] Take blank tobacco leaf samples, add Yiyadan to the final concentration of 5, 10, and 20 mg / kg, respectively, take test strips for detection, and repeat the measurement three times for each sample.
[0098] When testing tobacco leaf samples with test strips, when the added concentration of Yiyadan is 5 mg / kg, the test strip shows that the T line color is darker than the C line color or is consistent with the C line color, which is negative; When the added concentration of Yiyadan is 10 or 20 mg / kg, the test strip shows that the T-line color is lighter than the C-line color or the T-line has no color, which is positive, indicating that the test strip has no effect on tobacco leaves. The detection limit of Yiyadan is 10 mg / kg.
[0099] 2. False positive rate and false negative rate test
[0100] Take 20 copies of the positive samples of tobacco leaves with the known content of Yiyadan great...
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