4.7 Article

Dual electrophoretically-mediated microanalysis in multiple injection mode for the simultaneous determination of acetylcholinesterase and α-glucosidase activity applied to selected polyphenols

期刊

TALANTA
卷 224, 期 -, 页码 -

出版社

ELSEVIER
DOI: 10.1016/j.talanta.2020.121773

关键词

Acetylcholinesterase; alpha-glucosidase; EMMA; Phenolics

资金

  1. CNPq [444061/2018]
  2. FAPESC [2019TR0847]
  3. CAPES
  4. UFSC
  5. INCT-Catalise

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A novel method for simultaneous determination of AChE and α-glu activity was proposed, with reliable results and adherence to principles of green chemistry. Enzymatic kinetic parameters were tested and phenolic compounds were evaluated, showing promising inhibitory activity for both enzymes.
Acetylcholinesterase (AChE) and alpha-glucosidase (alpha-glu) are key target enzymes in the search for novel strategies in the treatment of Alzheimer's disease and type II diabetes. Therefore, methods to assess the enzyme inhibition are of great value in the research field. Here is proposed a novel a dual electrophoretically-mediated microanalysis for the simultaneous determination of both enzymes' activity. In order to do so, the various solutions required for both assays were introduced in the capillary electrophoresis system using the multiple injections approach. Enzymatic kinetic parameters were tested, K-m for AChE and alpha-glu were 3.81 and 0.43 mmol L-1. K-i values were 4.27 mu mol L-1 for neostigmine (an AChE inhibitor) and 0.40 mmol L-1 for acarbose (an alpha-glu inhibitor). Results of IC50 (concentration for 50% of inhibition) were 5.11 +/- 0.47 mu mol L-1 and 0.58 +/- 0,02 mmol L-1 for neostigmine and acarbose, respectively. All parameters (except for K-iY of neostigmine) were comparable with the literature, indicating a good reliability of the proposed method to evaluate these enzymes activity. Total time analysis was approximately 10 min, being possible to perform around 12 enzymatic assays per hour, with low sample and reagent consumption, thus satisfying some of the principles of green chemistry. The method was applied to evaluate 10 phenolic compounds, of wich p-coumaric acid showed the best inhibitory activity for AChE (40.14 +/- 4.75% at 10 mg L-1); and quercetin for alpha-glu (46.53 +/- 4.90% at 10 mg L-1).

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