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URL: https://dx.doi.org/DOI:10.17504/protocols.io.s9meh46

Authors: Maira Rubi, Maira Rubi Segura Campos

Summary: The assay was carried out following the protocol reported by Dineshkumar et al. [8]. Starch (2 mg) was suspended in a tube containing 0.2 mL of 0.5 M Tris-HCl (Sigma-Aldrich, USA) buffer (pH 6.9) with 0.01 M calcium chloride as substrate. The tube was boiled for 5 min and then preincubated at 37 °C for 5 min. Plant aqueous extract (1 mg) was dissolved with 1 mL of 0.1% of dimethyl sulfoxide in order to obtain a concentration of 1,000 µg/mL; then 0.2 mL of aqueous extract was added to the tube containing the substrate solution, 0.1 mL of porcine pancreatic amylase in Tris-HCl buffer (2 U/mL) was also added, and incubated for 10 min at 37 °C. Finally, the reaction was stopped with 0.5 mL of acetic acid (50% v/v) and centrifuged 5 min at 1,811 × g and 4 °C. The assay was performed in triplicate. The a-amylase inhibitory activity was calculated using the formula (Ac+) – (Ac-) – (As - Ab)/(Ac+) – (Ac-) × 100, where Ac+, Ac-, As, Ab are defined as the absorbance (595 nm) of 100% enzyme activity (only solvent with enzyme), 0% enzyme activity (only solvent without enzyme), test sample (with enzyme), and a blank (a test sample without enzyme), respectively.

Affiliations: , Universidad Autónoma de Yucatán

Version: 1

Publication Date: 2018

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