Methyl paraoxon fluorescence-colorimetric analysis method based on gold nanocluster-manganese dioxide nanosheet
A technology of methyl paraoxon and gold nanoclusters is applied in the field of biosensors, which can solve the problems of complex operation and low sensitivity, and achieve the effect of great application prospects.
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Embodiment 1
[0028] Example 1: Preparation of gold nanocluster AuNCs functionalized with bovine serum albumin
[0029] 5 mL of HAuCl 4 (10mmol L -1 ) and 5mL of BSA (50mg mL -1 ) and incubated at 37°C for 10 minutes, mixed the two, and stirred vigorously for 5 minutes. 0.5mL of NaOH (1mol L -1 ) into the above solution, stirred vigorously at 37°C for 12 hours to obtain a yellow AuNCs solution, purified by dialysis (1kDa), freeze-dried to obtain yellow AuNCs powder, dissolved and diluted to 0.20mg mL with ultrapure water -1 spare.
Embodiment 2
[0030] Example 2: AuNCs-MnO 2 Preparation of nanosheet composites
[0031] Take 2mL of the AuNCs solution of Example 1 and add it into 7.7mL of ultrapure water, and stir slowly for 5min. Then, 0.2mL MnCl 2 4H 2 O (50mmol L -1 ) was added to the above solution and stirred slowly for 1 hour; then 0.2mL NaOH (1molL -1 ), stirred slowly for 4 hours to obtain brown-yellow AuNCs-MnO 2 The composite material solution was purified by dialysis (1 kDa) and kept at 4°C for use.
Embodiment 3
[0032] Embodiment 3: Fluorescence-colorimetric sensing design of acetylcholinesterase
[0033] Detection of AuNCs-MnO 2 The sensitivity of the composite fluorescent system to AChE, 2mmol L -1 ATch and different concentrations of AChE (0 ~ 1000mU mL -1 ) and mix well, mix and react at 37°C for 25min, then add 50μL of the AuNCs-MnO 2 Mix well, and mix and react at 37°C for 15 minutes. As the concentration of AChE increased, the AuNCs-MnO 2 The fluorescence intensity gradually increased ( image 3 ), the absorbed light gradually decreases ( Figure 5 ), the color of the system gradually becomes lighter ( Image 6 ). Fluorescence recovery rate ((F–F 0 ) / F 0 , where F and F 0 Indicates the fluorescence intensity when AChE exists and does not exist) and the concentration of AChE is 1-80mU mL -1 Good linear relationship over the range (R 2 =0.999)( Figure 4 ).
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