Defender Roberto Cereceda was dropped from Chile's World Cup squad on Tuesday as coach Marcelo Bielsa finalised his 23-man party for South Africa.
Bielsa named a provisional 30-man squad last month and then whittled it down, first to 25 and then 24, meaning he had to drop one more player before the FIFA deadline on Tuesday.
Left wing-back Cereceda, who plays for Chile's leading side Colo Colo, was the unlucky player to miss out, despite having captained his country in a friendly as recently as May 5.
Chile leave for South Africa on Saturday and play their opening group H match against Honduras on June 16.
Goalkeepers: Claudio Bravo (Real Sociedad), Miguel Pinto (Universidad de Chile), Luis Marin (Union Espanola)
Defenders: Pablo Contreras (PAOK Athens), Ismael Fuentes (Universidad Catolica), Mauricio Isla (Udinese), Gonzalo Jara (West Bromwich Albion), Gary Medel (Boca Juniors), Waldo Ponce (Universidad Catolica), Arturo Vidal (Bayer Leverkusen).
Midfielders: Rodrigo Tello (Besiktas), Gonzalo Fierro (Flamengo), Carlos Carmona (Reggina), Marco Estrada (Universidad de Chile), Rodrigo Millar (Colo Colo), Matias Fernandez (Sporting Lisbon), Jorge Valdivia (Al Ain).
Forwards: Mark Gonzalez (CSKA Moscow), Esteban Paredes (Colo Colo), Juan Beausejour (America), Fabian Orellana (Xeres), Alexis Sanchez (Udinese), Humberto Suazo (Real Zaragoza).