Acute and sub-acute toxicological evaluation of a bioactive compound mixture in healthy Wistar rats

The increasing interest in bioactive compounds necessitates a thorough understanding of their <i>in vivo</i> safety profiles before they are developed as therapeutic drug leads. The present study aimed to evaluate acute and sub-acute toxicological effects of a compound mixture composed of garcinol, piperine, butyl oleate, pipnoohine, and bismurrayanimbine, combined in a molar mass ratio of 9:33:1:4:1 at the doses of 10 mg kg^-1, 25 mg kg^-1, and 50 mg kg^-1 in healthy Wistar rats, following Organization for Economic Co-operation and Development guidelines. A single oral dose of the compound mixture (10 mg kg^-1, 25 mg kg^-1, and 50 mg kg^-1) was administered, and the rats were closely observed over a subsequent 14-day period. Further, the compound mixture was administered orally at the same three doses to Wistar rats continuously for 28 days. The compound mixture at the three doses did not produce mortality or abnormal behavioral changes throughout the 14 days. No significant alterations in hematological parameters or biochemical markers such as alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and creatinine (p > 0.05) were observed. Histopathological analysis revealed no treatment-related changes in the hematoxylin and eosin-stained sections of the liver, kidney, heart, spleen, stomach, small intestine, and lung tissues. In conclusion, the oral administration of compound mixture at 10 mg kg^-1, 25 mg kg^-1, and 50 mg kg^-1 for 28 days was found to be safe in healthy Wistar rats via biochemical (liver enzymes, kidney function tests), hematological (full blood count analysis), and histological assessments of hematoxylin and eosin-stained tissue sections.