On-site quantitative detection platform for organophosphorus pesticides integrated with target-responsive hydrogel and smartphone
A technology of organophosphorus pesticides and smartphones, applied in the field of biosensors, can solve the problems of pesticide concentration requirements, low detection sensitivity, cumbersome test methods, etc., and achieve the effects of fast response, simple construction process, and low cost
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Embodiment 1
[0029] Example 1: MnO 2 Synthesis of NFs hydrogels
[0030] First add 0.5mg mL -1 BSA with 5.0 mL MnCl 2 (4.0×10 -3 mol L -1 ) aqueous solution was mixed, and the mixture solution was continuously stirred at room temperature for 1 hour. Subsequently, with 1.0mol L -1 NaOH solution adjusted the pH of the reaction mixture to 10.0. After the reaction, it was allowed to stand for 4 hours, and the product was stored at 4.0°C. Then, fully dissolve SA in Tris-HCl buffer pH 8.0, quickly transfer SA (20mg mL -1 ) into a 96-well plate. Next add 0.025mg mL -1 MnO 2 NFs were added to SA, and immediately after mixing well, CaCl was added dropwise to the above mixture 2 (5.0mg mL -1 ), made of uniform BSA-MnO 2 NFs-SA hybrid hydrogel;
Embodiment 2
[0031] Embodiment 2: the making of kit
[0032] 3.0 μg mL -1 Paraoxon aqueous solution (20μL) and acetylcholinesterase (8.0U mL -1 ) aqueous solution (20 μL) was mixed and incubated at 37° C. for 20 minutes. Then add 40 μL acetylcholine (100 mmol L -1 ) and choline oxidase (4.0U mL -1 ) aqueous solution. After incubating at 37°C for 30 minutes, the reaction solution was added to the BSA-MnO obtained in Example 1 2 NFs-SA mixed hydrogel, thereby obtaining the MnO described in the present invention 2 NFs Hydrogel Kit. Add TMB (1.5mmol L -1 , 20 μL) chromogenic reagent, the color reaction of the kit is triggered. At the concentration of paraoxon in this example, the color of the kit is dark blue with a gray value of 79.405.
Embodiment 3
[0033] Embodiment 3: the application of kit to OPs detection
[0034] According to the method of Example 2, different concentrations of OPs (0, 0.0005, 0.005, 0.05, 0.3, 0.5, 1.5 and 3.0 μg mL -1 , the corresponding gray values at different concentrations are 159.07, 119.81, 109.33, 90.95, 84.79, 82.36, 81.42, 79.405) MnO 2 NFs hydrogel kit, and then drop 20μL, 1.5mmol L -1 After TMB chromogen, the color of the kit can change from colorless to dark blue as the concentration of OPs increases with the naked eye, and the color change is recorded using an application (APP) made by a smart phone. It has a built-in image processing program, which can convert the corresponding image information into gray value, and further display the linear relationship with the logarithm of paraoxon concentration, so as to realize the on-site handheld detection of OPs (such as figure 2 shown);
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