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A colorimetric assay for acetylcholinesterase activity and inhibitor screening based on the thiocholine–induced inhibition of the oxidative power of MnO2 nanosheets on 3,3′,5,5′–tetramethylbenzidine

Microchimica Acta volume 185, Article number: 446 (2018) Cite this article

Abstract

The authors describe a colorimetric method for the determination of the activity of acetylcholinesterase (AChE). Manganese dioxide (MnO2) nanosheets directly reacts with 3,3′,5,5′–tetramethylbenzidine (TMB) in the absence of hydrogen peroxide (H2O2). This leads to the formation of a blue product (oxTMB) with an absorption peak at 652 nm. If AChE hydrolyzes its substrate acetylthiocholine chloride, thiocholine is formed which blocks the oxidative power of the MnO2 nanosheets. Hence, oxTMB will not be formed. The decreased absorbance is directly related to the AChE activity in the 0.01–1.0 mU·mL−1 range. The detection limit is 0.01 mU·mL−1 and the relative standard deviation is 1.2% (for n = 11 at 0.5 mU·mL−1). The method was also applied to screen for inhibitors of AChE.

Based on the oxidizing properties of manganese dioxide nanosheets (MnO2 nanosheets), we report a colorimetric method for determining acetylcholinesterase activity with the chromogenic substrate 3,3′,5,5′-tetramethylbenzidine (TMB).

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Acknowledgements

The authors gratefully acknowledge financial support from Fundamental Research Funds for the Central Universities (Grant No. xjj2015121).

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  1. Department of Chemistry, School of Science, Xi’an Jiaotong University, Xi’an, 710049, China

    Yuan Sun, Haonan Tan & Yinhuan Li

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  1. Yuan Sun
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Correspondence to Yinhuan Li.

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Sun, Y., Tan, H. & Li, Y. A colorimetric assay for acetylcholinesterase activity and inhibitor screening based on the thiocholine–induced inhibition of the oxidative power of MnO2 nanosheets on 3,3′,5,5′–tetramethylbenzidine. Microchim Acta 185, 446 (2018). https://doi.org/10.1007/s00604-018-2974-4

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Keywords

  • Acetylcholinesterase
  • Acetylcholinesterase activity
  • Colorimetry
  • MnO2 nanosheets
  • 3,3′,5,5′–Tetramethylbenzidine
  • Oxidation
  • Acetylthiocholine chloride
  • Inhibitor screening
  • Neostigmine