The dietary fiber extracted from cassava pulp, composed of crude fiber, neutral detergent fiber (NDF), and cellulose content, demonstrates promise as a functional food ingredient. The study’s objectives encompassed the assessment of short-term toxicity and the evaluation of its potential cholesterol-lowering effects. The results indicated that cassava pulp dietary fiber (CDF) is well-tolerated with non-toxic thresholds determined at 10.01 g/kg body weight/day for male rats and 11.21 g/kg body weight/day for female rats during the short-term toxicity assessment. Furthermore, CDF exhibited notable cholesterol-lowering effects, significantly reducing serum triglyceride and serum total cholesterol levels, along with decreased liver total lipids and liver cholesterol levels. In contrast, it led to significant increases in fecal total lipids and cholesterol when compared to the control group. Most notably, there were no significant differences in terms of serum triglyceride, serum total cholesterol, liver total lipids, and liver cholesterol between CDF and the conventional cholesterol-lowering medication, simvastatin. These findings underscore the potential of cassava pulp dietary fiber as a natural and safe alternative for managing hyperlipidemia and related conditions. It offers a valuable avenue for the development of functional foods aimed at improving cardiovascular health and further investigation for its potential application in the field of nutraceuticals. © 2023 by the authors.