2.6. Acute Toxicity Test. R. nasutu

2.6. Acute Toxicity Test. R. nasutus (50–250mg/kg body weight) was orally administered to rats for acute toxicity stud- ies. Each group was observed individually for signs of toxicity and behavioral changes such as hyperactivity, grooming, con- vulsions, sedation, or hypothermia. These observations began 1 h after dosing and were continued at least once daily for 14 days. The mortality rate was also calculated.
2.7. Biochemical Measurements. At the end of the study (30 days), after an overnight fasting, the animals were sacrificed by cervical dislocation following anesthesia using isoflurane. The liver tissue was excised and washed with ice-cold saline and was immediately immersed in liquid nitrogen and stored at −80∘C for further biochemical analysis. Then, the measure- ments of liver enzyme activity and biochemical assays were performed.
AST and ALT activities were assayed using the method of Reitman and Frankel [28]. The total carbohydrate content was estimated based on the method established by Carroll et al. [29]. Glycogen content was determined as described by Saifter et al. [30]. The protein content was estimated by the method of Lowry et al. [31] with slight modifications. All enzymatic assays in this study were performed using crude liver homogenate.
2.8. Statistical Analysis. The results were expressed as the mean ± SD (