2.5. Acute Toxicity Assay

The acute toxicity assay followed the guidelines of the Organization for Economic Co-operation and Development (OCDE)-423/2001 [27]. This protocol states that, when previous studies have assessed the acute toxicity of a known sample, the initial dose used can be the maximum dose without side effects. As previously reported by Sachetti et al. [25], C. reticulata oleoresin was tested in a single dose of 2000 mg/kg. We used twelve 9-week-old female rats weighing 150 to 180 g. Six rats received the oleoresin and six were kept as the control group, receiving the same dose (2000 mg/kg) of water. The acute toxicity assay was performed with three rats per group (control and treated) and repeated after 14 days with another three rats per group, totaling twelve rats. Several parameters such as general activity, irritability, contortion, ataxia, tremors, convulsions, piloerection, hypothermia, breathing, cyanosis, hyperemia, and death were analyzed.