Simultaneous estimation of epigallocatechin, fisetin, and quercetin in Acacia suma and its potential against postprandial hyperglycemia

Abstract

Anti-diabetic activity of the plants may follow one or the other mechanism such as their ability to restore the pancreatic tissue function or increase in insulin output or inhibiting the intestinal absorption of glucose or inhibiting the pancreatic enzymes like alpha-amylase, alpha-glucosidase, etc. The alpha-amylase and alpha-glucosidase inhibition assays were performed to analyze the anti-diabetic potential of Acacia suma. RP-HPLC method was developed and validated for simultaneous estimation of Epigallocatechin (EGC), Fisetin (FT), and Quercetin (QT) in hydroalcoholic extract (HAE) of Acacia suma and formulations. The simultaneous estimation was performed using the Luna C18 column with 20µl sample injection volume. Whereas, acetonitrile and phosphate buffer (0.1%, 30:70 v/v, pH 2.7) was used for resolution as mobile phase with column temperature 40 ^OC and flow rate 1 ml/min. The IC₅₀ for alpha-amylase inhibition and alpha-glucosidase inhibition activity was found to be 12.5µg/ml and 93.79µg/ml respectively was compared with standards. The developed method was found simple, specific, precise and linear with regression coefficient (R² = 0.999) over the selected range of concentration (2.5-160 μg/mL) having detection limits 0.076, 0.012, 0.010 μg/mL and quantification limits 0.231, 0.038, 0.031 μg/mL for EGC, FT and QT respectively. In-vitro alpha-amylase and alpha-glucosidase inhibition assay support the traditional claim of the plant in the treatment of metabolic disorders. The established RP-HPLC method demonstrates the precise and easy determination of EGC, FT, and QT and is effectively studied by a gradient elution system.