The Capitals captain had a 39 second shift early in the game and then left the team bench. The team later announced that he would not return due to a lower body injury.
Ovechkin has out since April 22 after suffering a lower-body injury against the New York Islanders.
Defenceman John Carlson also returned to the lineup Monday night against the Rangers. Carlson missed the last two games with a lower-body injury.
Evgeny Kuznetsov and Ilya Samsonov will miss Monday's game for disciplinary reasons while T.J. Oshie for a personal family matter, according to The Washington Post's Samantha Pell.
Ovechkin has 24 goals and 42 points in 43 games this season while Carlson has 10 goals and 42 points in 49 games. The Capitals sit second in the East Division trailing the Pittsburgh Penguins by two points.