3.7. In Vivo Acute Oral Toxicity Study and Gross Behavioral Studies

The in vivo acute oral toxicity study for the newly synthesized scaffolds 4b and 4h was performed according to the OECD Guideline no. 425 [60] using Swiss albino mice (18–22 gm weight) quarantined at the animal house at Y.B. Chavan College of Pharmacy, Aurangabad IAEC approval number CPCSEA/IAEC/P’col-52/2015-16/115. Each group consisted of six mice (overnight fasted) and kept in a colony cage at 25 ± 2 °C with 55% relative humidity and 12 h of light and dark cycle. A specified dose of 100, 250, 500, 750, 1000, 1500 and 2000 mg/kg body weight of mice was administered orally as a single dose. The acute toxic symptoms and the behavioral changes produced by the test compounds were observed continuously for 4 h periods at 8th, 12th and 24th h. Onset of toxic symptoms and the gross behavioral changes were also recorded. These animals were maintained for further 10 days with observations made daily. In case the animal appeared moribund (dying) the animal was sacrificed in a humane way and it is considered to have died because of toxicity.