Optimisation and application of antihemolytic assay on Fagopyri herba extracts

Abstract

Fagopyri herba, a herbal drug derived from common buckwheat (Fagopyrum esculentum Moench) is a very rich source of various phenolic compounds which are known as potential antioxidants. The main active compound is a flavonol glycoside rutin. The aim of this study was to investigate antihemolytic activity of the Fagopyri herba extracts as a parameter which indicates their antioxidant potential in biological systems. Antihemolytic activity assay measures the ability of a tested extract to inhibit hemolysis of erythrocytes in buffer solution caused by oxidative damage of their cell membrane. Since this test is highly dependent on various parameters (cell suspension concentration, species from which the blood was taken, concentration of reagents, type of extract), they were optimized for testing the investigated water and ethanol Fagopyri herba extracts. Optimal concentration of hydrogen peroxide solution was found to be 0.0625% (v/v) while 2 μM of sodium azide was added to the erythrocyte suspension to inhibit catalase enzyme. Ethanolic extract showed lower IC50 value (3.46±0.185 mg/mL) in comparison to water extract (42.2±7.27 mg/mL), which is in accordance with their rutin content determined by high performance liquid chromatography.