Colorimetric method for detecting acid phosphatase or organophosphorus pesticide on basis of mimic biomimetic oxidase activity of manganese dioxide
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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
Pending Publication Date: 2020-06-26
NANJING MEDICAL UNIV
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[0006] The purpose of the present invention is to overcome the shortcomings of low sensitivity, time-consuming detection, high cost and cumbersome steps of the existing acid phosphatase detection method, and to provide a colorimetric sensing system based on the activity of manganese dioxide biosimulated oxidase
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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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Abstract
The invention discloses a colorimetric method for detecting acid phosphatase on the basis of mimic biomimetic oxidase activity of manganese dioxide. The colorimetric method for detecting the acid phosphatase on the basis of the mimic biomimetic oxidase activity of the manganese dioxide comprises the following steps: producing ACP solutions and an AAP solution by adopting a NaAc-HAc buffer solution, and mixing the multiple ACP solutions with different concentrations, the AAP solution, MnO2 nanosheets and ABTS; measuring ultraviolet absorbancy degrees A and A0 at the wavelength of 420 nm to obtain a difference value deltaA of the absorbancy degrees, so as to obtain a linear relation between the ACP concentrations and the difference value deltaA of the absorbancy degrees; and calculating a concentration of the ACP of a to-be-detected sample according to the linear relation and the difference value deltaA of the absorbancy degrees of the to-be-detected sample. A colorimetric method for detecting an organophosphorus pesticide on the basis of the mimic biomimetic oxidase activity of the manganese dioxide is further disclosed; and the organophosphorus pesticide is added on the basis of the colorimetric method for detecting the acid phosphatase. According to the colorimetric method for detecting the acid phosphatase on the basis of the mimic biomimetic oxidase activity of the manganesedioxide and the colorimetric method for detecting the organophosphorus pesticide on the basis of the mimic biomimetic oxidase activity of the manganese dioxide, color comparison is conducted on the acid phosphatase and the organophosphorus pesticide, and the colorimetric method for detecting the acid phosphatase on the basis of the mimic biomimetic oxidase activity of the manganese dioxide and the colorimetric method for detecting the organophosphorus pesticide on the basis of the mimic biomimetic oxidase activity of the manganese dioxide have the advantages of being simple, high in sensitivity, low in cost and high in detection throughput.
Description
technical field [0001] The invention belongs to the technical field of analysis and detection, and relates to a colorimetric detection method for acid phosphatase or organophosphorus pesticide based on manganese dioxide imitation bioimitation oxidase activity, more specifically, relates to a biomimetic oxidation method based on manganese dioxide Enzyme activity, used for the detection of acid phosphatase and organophosphorus pesticides. Background technique [0002] Acid phosphatase (ACP) is a ubiquitous multifunctional enzyme mainly distributed in the blood and tissues of mammals, which can hydrolyze the phosphate bonds of organophosphorus compounds under weakly acidic conditions. As a phosphomonoesterase, acid phosphatase plays an important role in regulating various physiological changes in cells. At the same time, a large number of studies have pointed out that the abnormal expression and activation of ACP is associated with many human diseases, including hyperparathyro...
Claims
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