The total phenolic content of E. hirta parts extracts was
determined by the Folin-Ciocalteu reagent according to
Singleton and Rossi procedure[12]. 200 毺L plant extract
(three replicates of 1.0 mg/mL) were introduced into test
tubes; 500 毺L of 10% Folin-Ciocalteu’s reagent, 500 毺L
of distilled water and 800 毺L of 7.5% saturated aqueous
sodium carbonate (Na2CO3) were added. The tubes were
mixed thoroughly and allowed to stand in dark condition at
ambient temperature for 30 min. Absorption was measured
at 765 nm using spectrophotometer (Hitachi U-1900,
Tokyo, Japan). Distilled water was used as a blank and
gallic acid (0-250 mg/L) was used to produce standard