Erythrocyte lysis assay

The cytotoxicity of 3BP on human erythrocyte cells was analyzed using the protocol described previously by Turecka et al., [31]. Briefly, 5 mL of human blood was collected from healthy volunteers and transferred to a fresh 15 mL falcon tube containing an anticoagulant agent (sodium citrate). The total erythrocyte cells were harvested by centrifugation at 3000 rpm for 5 min and dissolved in PBS at the final concentration of 2%. Further, the blood suspension was exposed to different concentrations (0–1024μg/mL) of 3BP at 37°C for 1 h. Subsequent to incubation, the supernatant was collected to spectroscopically measure the erythrolytic activity at 415 nm.