Anti-inflammatory activity

Anti-inflammatory in vitro activity was evaluated using HRBC membrane stabilization method (Emamuzo et al. 2010). Blood sample collected from healthy volunteers was mixed with equal volume of sterilized Alsevers solution and centrifuged at 3000 rpm to separate packed cell volume (PCV). The separated PCV was washed with iso-saline solution and a 10% (v/v) suspension was prepared in iso-saline. Different EO concentrations were separately mixed with 1 mL of phosphate buffer, 2 mL of hypo-saline, and 0.5 mL of HRBC suspension. The assay mixtures were incubated at 37 °C for 30 min and centrifuged at 3000 rpm. The isolated hemoglobin content was estimated by a spectrophotometrically (560 nm). The percentage hemolysis was estimated by assuming the hemolysis produced in the control as 100%, which was calculated as: percentage protection = 100 − (OD_sample/OD_control) × 100.