4.3.6. Cytotoxic Assay Using Brine Shrimp Lethality Assay (BSLA)

The cytotoxic effects of B. Pachyancantha leaf extract and BPL@NiONPs were evaluated using brine shrimp cytotoxicity assay. Artificial sea water was prepared by adding 3.8 g sea salt in 1 L distilled water using a partitioned hatching chamber. Eggs of brine shrimp (Artemia salina) were put in the covered portion of the chamber and incubated for 48 h at 30 °C. Four different concentrations of BPL extract and BPL@NiONPs (200–1 µg/mL) were added in each vial and their final volume was adjusted up to 5 mL by adding sea water. Then, ten mature brine shrimps were introduced into each vial and left under the lamp. After 24 h, alive brine shrimps were counted [58]. Potassium permanganate was used as a positive control while sea water and distal water were used as a negative control. Lethality concentration (LC₅₀) values and percentage mortality were calculated.