α-glucosidase inhibition assay

Hasnain Jan; Hazrat Usman; Muzamil Shah; Gouhar Zaman; Sadaf Mushtaq; Samantha Drouet; Christophe Hano; Bilal Haider Abbasi

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α-glucosidase inhibition assay

HJ Hasnain Jan

HU Hazrat Usman

MS Muzamil Shah

GZ Gouhar Zaman

SM Sadaf Mushtaq

SD Samantha Drouet

CH Christophe Hano

BA Bilal Haider Abbasi

This method is extracted from research article: BMC Complement Med Ther, Jun 2021

Phytochemical analysis and versatile in vitro evaluation of antimicrobial, cytotoxic and enzyme inhibition potential of different extracts of traditionally used Aquilegia pubiflora Wall. Ex Royle

DOI: 10.1186/s12906-021-03333-y

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The anti-diabetic potential of extracts was further determined by α-glucosidase inhibition bioassay using a previously reported protocol with minute modifications [42, 43]. In the experiment, 50 mL of phosphate buffer (pH 6.8) supplemented with 100-mg BSA (bovine serum albumin) was used to dissolve α-glucosidase (Saccharomyces cerevisiae, Sigma-Aldrich). Reaction mixtures constituting 10 μL of tested sample, phosphate buffer (490 μL; pH 6.8) and p-nitrophenyl α-D-glucopyranoside (5 mM; 250 μL) were kept sepatately for incubation at 37 °C for 5 min. Two hundred fifty microlitre α-glucosidase (0.15 unit/mL) was then added to each mixture followed by incubation for 15 min at 37 °C. After terminating reaction, by adding 2 mL Na₂CO₃ (200 mM) solution, absorption was recorded using a UV-Vis spectrophotometer at 400 nm. The assay is based on the quantification of p-nitrophenol release from p-nitrophenyl α-D-glucopyranoside. In the experiment, acarbose was employed as a positive control and assay was repeated three times.

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