Ola Aina and Mikel Agu were the last two casualties as Nigeria coach Gernot Rohr settled on his final 23-man World Cup squad.
Neither player took part in the 2-1 friendly defeat to England at Wembley, where the Super Eagles recovered from a dismal first half to provide some confidence going into the tournament in Russia.
"It's Ola Aina and Mikel Agu," Rohr told KweséESPN just before the team made their way to the airport on Sunday morning to leave for the final phase of their training camp in Austria.
"They will be on standby. We go with 23 players to Austria; if there is another problem or injury like happened four years ago when Elderson was injured, they will be on standby for that."
Aina was in competition with Bryan Idowu and Elderson Echiejile for one left-back spot, while Agu had Joel Obi and John Ogun to contend with in midfield.
Idowu's performances appear to have established him as the first-choice for Rohr, while Echiejile's experience gave him an additional edge.
With Agu, he appears to have lost out to Joel Obi's ability to slot into different positions in the line-up. The former Inter Milan man can play as left-back, left midfielder, central midfielder and even left forward.
Nigeria play their last official friendly match against Czech Republic on June 6 before heading out to Russia where they will face Croatia, Iceland and Argentina in the Group stages.
Goalkeepers: Francis Uzoho (Deportivo La Coruna), Ikechukwu Ezenwa (Enyimba), Daniel Akpeyi (Chippa United).
Defenders: William Troost-Ekong and Abdullahi Shehu (Bursaspor), Tyronne Ebuehi (Ado Den Haag), Elderson Echiejile (Cercle Brugge KSV), Bryan Idowu (Amkar Perm), Chidozie Awaziem (Nantes FC), Leon Balogun (Brighton), Kenneth Omeruo (Kasimpasa).
Midfielders: Mikel John Obi (Tianjin Teda), Ogenyi Onazi (Trabzonspor), Wilfred Ndidi (Leicester City), Oghenekaro Etebo (Las Palmas), John Ogu (Hapoel Be'er Sheva), Joel Obi (Torino, Italy).
Forwards: Ahmed Musa (CSKA Moscow), Kelechi Iheanacho (Leicester City), Victor Moses (Chelsea), Odion Ighalo (Changchun Yatai), Alex Iwobi (Arsenal), Simeon Nwankwo (Crotone).