2.3. Cytotoxicity test (Brine shrimp lethality assay)

Brine shrimp lethality bioassay (Krishnaraju et al., 2006) was used to determine the cytotoxic capability of the P. wallichiana stem, D. oloeidies fruit and B. chinensis (whole plant) extracts using Artemia salina (test organism).The Fig. 2 represents the entire assay used in this experiment. Artificial sea water was composed of commercial salt mixed with double distilled under continuous aeration for 48 h. Brine shrimp eggs were hatched in conical shaped vessel (1L) filled with sterile artificial seawater. After hatching the active nauplii were collected for the test. Then stock solution was prepared by dissolving 10 mg of plant extract in 1 mL of distilled water. Concentrations prepared by serial dilution from the stock solution (10, 100 and 1000 μg/ml) were added to the pre-labelled test tubes. Then from each concentration 1 mL solution was added into the test tubes having 1 mL of seawater and 30 nauplii. The mortality rate was evaluated after 24 h by measuring the number of dead nauplii. The Etoposide, a standard drug was used as positive control.

Brine Shrimp Lethality Assay of plant extracts.