Optimization and Validation of a Microscale Method to Assess  α-Glucosidase Inhibition Activity

Overview

Journal Curr Anal Chem

Publisher Bentham Science Publishers

Specialty Chemistry

Date 2018 Oct 9

PMID 30294249

Citations 3

Authors

Graciela Granados-Guzman

Rocio Castro-Rios

Noemi Waksman de Torres

Ricardo Salazar-Aranda

Affiliations

Soon will be listed here.

Abstract

Background: Microscale in vitro assays are fast, simple, and inexpensive, with reduced reagent quantities, waste, and experimental animal use. However, they have low reproducibility and low correlation with the results of in vivo models, possibly due to differences in precision and accuracy in methodologies between laboratories.

Objective: The objective was the optimization and validation of an in vitro assay, carried out on microscale, to assess the inhibition of α-glucosidase activity, which is indicative of antihyperglycemic activity.

Methods: The optimization was carried out using a fractional factorial design taking into account the best inhibition percentage and the absorbance of the controls. With the optimized experimental conditions in hand, we carried out method validation.

Results: The optimized conditions were as follows: enzyme concentration, 0.55 U/mL; substrate concentration, 111.5 μM; and 17.5 min incubation at 37°C. A linear range between 100 and 310.2 μg/mL of acarbose (r2 0.994) was established. The RSD was <2% and the % error was <3%. The Z factor was >0.96. This method was applied to four plant extracts, one of which was found to be very active.

Conclusion: The method was found to be accurate, precise, selective, linear, and reliable in evaluating the antihyperglycemic activity of natural extracts in vitro.

Citing Articles

Green solvent-based extraction of three Fabaceae species: A potential antioxidant, anti-diabetic, and anti-leishmanial agents.

Fatima I, Safdar N, Akhtar W, Ayaz A, Ali S, Elansary H Heliyon. 2024; 10(13):e33668.

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Discovery of novel flavonoid derivatives as potential dual inhibitors against α-glucosidase and α-amylase: virtual screening, synthesis, and biological evaluation.

Mai T, Phan M, Thai T, Lam T, Lai N, Nguyen T Mol Divers. 2023; 28(3):1629-1650.

PMID: 37369956 DOI: 10.1007/s11030-023-10680-0.

Screening natural product extracts for potential enzyme inhibitors: protocols, and the standardisation of the usage of blanks in α-amylase, α-glucosidase and lipase assays.

Lankatillake C, Luo S, Flavel M, Lenon G, Gill H, Huynh T Plant Methods. 2021; 17(1):3.

PMID: 33407662 PMC: 7789656. DOI: 10.1186/s13007-020-00702-5.

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Optimization and Validation of a Microscale Method to Assess α-Glucosidase Inhibition Activity

Optimization and Validation of a Microscale Method to Assess  α-Glucosidase Inhibition Activity