The present investigation evaluated the safety of the methanol extract from the fruit body of Phellinus pini Ames (PPA) by determining its potential toxicity after a subacute administration in rats. The extract was orally administered in doses of 1 g/kg, 2 g/kg, and 4 g/kg daily for 14 days to rats. Body weight, biochemical, and hematological parameters were determined at the end of 14 days of daily administration. The no-observed adverse effect levels (NOAEL) of the extract were 4 g/kg, when given by gavage routes. Daily oral administration of PPA extract for up to 14 days did not result in the death of significant changes in the body weight, hematological, and mainly biological parameters. In biological analysis, some significant changes occurred, including triglyceride and blood urea nitrogen (BUN), indicating that the PPA extract has liver and kidney-modulating activity. The PPA extract was found to be low or non-toxic in rats.