Teenage Mutant Ninja Turtles: Tournament Fighters is a fighting game in the tradition of the Mortal Kombat or Street Fighter series, pitting the player against one opponent at a time. The game features eight fighters: the four turtles, Casey Jones, April O'Neil, Ray Fillet (from the comics) and the original character Sisphyus.

The gameplay is similar to other games in this genre: each character has standard attacks (normal and strong) as well as special attacks which require button combinations to perform. The goal is to deplete the other fighter's life energy completely. However, a losing opponent has one last straw: a very powerful attack which is only unlocked shortly before death. The amount of rounds can be set between one and seven.

The game's modes include a story mode, in which the selected hero has to battle against Krang's evil clones of the player characters in order to save the kidnapped Splinter, a two-player mode, a practice mode and a tournament mode which requires the player to fight against multiple opponents one after another with only one life bar.