Colorimetric method for detecting acid phosphatase or organophosphorus pesticide on basis of mimic biomimetic oxidase activity of manganese dioxide
An oxidase activity, organophosphorus pesticide technology, applied in the field of analysis and detection, can solve the problems of high cost, time-consuming detection, cumbersome steps and the like
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Embodiment 1
[0055] In order to obtain the optimal ACP detection environment, the inventor takes the absorbance difference ΔA as the standard ((ΔA=A 0 – A, A 0 and A represent the absorbance value at 420nm in the presence and absence of ACP, respectively), the influence factor MnO 2 The concentration of nanosheets in the system, the incubation time of AAP and ACP, the pH, and the concentration of AAP in the system were optimized.
[0056] (1), MnO 2 Effect of nanosheet concentration on ΔA
[0057] Step (a), using pure water (pH=7.0) to prepare ACP solution and AAP solution, mixing ACP solution and AAP solution, and incubating at 37°C for 10 minutes; without adding ACP as a control;
[0058] Step (b), MnO 2 The nanosheet solution is mixed with the mixed system obtained in step (a), and incubated at 37° C. for 10 minutes;
[0059] Step (c), add ABTS solution to the mixed system obtained in step (b), incubate at 37° C. for 50 min to obtain a colorimetric sensing system; in the colorimetric...
Embodiment 2
[0080] A colorimetric detection method for acid phosphatase based on manganese dioxide imitation bioimitation oxidase activity, comprising the following steps:
[0081] Step (a), adopt 0.2mol L -1 , pH 4.6 NaAc-HAc buffer solution to prepare different concentrations of ACP solution and AAP solution, mix different concentrations of ACP solution and AAP solution, and incubate at 37°C for 20min;
[0082] Step (b), MnO 2 The nanosheet solution is mixed with the mixed system obtained in step (a), and incubated at 37° C. for 10 minutes;
[0083] Step (c), adding the chromogenic substrate ABTS to the mixed system obtained in step (b), incubating the reaction at 37°C for 50 minutes to obtain a colorimetric sensing system; in the colorimetric sensing system, the concentration of ACP is 0.075 ,0.12,0.15,0.24,0.3,0.36,0.45,0.6,0.75,1.2,1.5,2.25,2.7,3mU·mL -1 , AAP concentration is 1mM, MnO 2 The concentration of nanosheets is 0.075 mg·mL -1 , the concentration of ABTS in the system ...
Embodiment 3
[0087] Selective experiment of detection method of the present invention
[0088] Replace the ACP in step (a) of Example 2 with 100 μM tyrosine, phenylalanine, lysine, sarcosine, aspartic acid, human serum albumin, bovine serum albumin, glucose, lysozyme trypsin, alkaline phosphatase, 1mMK + , Ca 2+ , Na + ,Mg 2+ ,Zn 2+ Etc. Interfering substances, repeat steps (b), (c), (d) in embodiment 2, other conditions are constant, detect absorbance difference, obtain the selectivity result of this method detection target ACP, see Figure 5 , it can be seen from the figure that the method of the present invention has good selectivity to acid phosphatase.
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