We studied the diet of Liophis reginae semilineatus (Wagler, 1824) from eastern Amazon, Brazil, based on the analysis of 182 preserved specimens. Thirty-six individuals had prey in their stomachs; 34 (95%) contained exclusively anurans and 2 (5%) contained both anurans and lizards. The most common prey items were small Leptodactylus sp. (33.3%), followed by Physalaemus ephippifer (Steindachner, 1864) (10.3%). Prey ingested head-first (78%; n = 25) were significantly larger than prey ingested tail-first (22%; n = 7). Females of L. reginae semilineatus have longer and wider heads than conspecific males with the same body length, which correspond to sexual divergences in the diet (size of the prey). No correlation was found between snake head length vs. prey size (SVL, width and mass). Liophis reginae semilineatus is an anurophagous snake that probably forages actively on the ground.