2.6. Acute toxicity assay

To allow for acclimatisation to the laboratory conditions, the rats were selected randomly, marked to allow individual recognition, and kept in their cages for at least 5 days prior to dosing. The animals were kept without food for overnight prior dosing but had access to water. Acute toxicity study was performed with a Limit test at 2000 mg/kg p.o. as single dose for all the extracts as shown in the Fig. 1. The dose was administered to the animal based on their body weight. The animals were closely observed for first 30 min, then for 4 h. Food was provided after 1–2 h of dosing. After survival of the treated animal 4 more animals were treated with the same dose.

Grouping of animals for acute toxicity studies.

A control group of rats (n = 5) were administered with 0.3% CMC in the same volume of that of the treated group. All the groups were closely observed for 6 h and then at a regular interval for 14 weeks. Survived animals were observed for other toxic effects. The weight of the animals was monitored from the beginning of the experiment and the blood samples were collected by cardiac puncture under anaesthesia with chloroform and serum was separated for biochemical and haematological evaluations. After the end of the experiment the animals were sacrificed by cervical dislocation and vital organs were excised, weighed and preserved in 10% formalin for histopathological evaluation.