Juventus talisman Ronaldo has not appeared at international level since the World Cup, with the national coach granting him an extended leave of absence.
Reports suggested he could return to face Italy and Poland, but Santos has honoured the joint decision not to consider the forward until 2019.
"The only thing I think about Ronaldo is that he should win the Ballon d'Or," Santos replied when asked about Portugal's all-time top scorer.
There are recalls for defenders Jose Fonte and Raphael Guerreiro, along with midfield pair Andre Gomes and Joao Mario.
Portugal need either a point or a win against Italy to finish top of Group 3 in Nations League A.
Santos' side beat the Italians 1-0 at home back in September, before picking up a 3-2 away win against Poland the following month and will be favourites to progress as group winners.
PORTUGAL SQUAD IN FULL:
Goalkeepers: Beto (Goztepe), Rui Patricio (Wolves), Claudio Ramos (Tondela)
Defenders: Joao Cancelo (Juventus), Ruben Dias (Benfica), Jose Fonte (Lille), Raphael Guerreiro (Borussia Dortmund), Luis Neto (Zenit), Pepe (Besiktas), Mario Rui (Naples), Cedric Soares (Southampton)
Midfielders: William Carvalho (Real Betis), Bruno Fernandes (Sporting), Andre Gomes (Everton), Joao Mario (Inter), Ruben Neves (Wolverhampton), Danilo Pereira (Porto), Pizzi (Benfica), Renato Sanches (Bayern Munich)