Thiago, Koke and Inigo Martinez are all included as the likes of Xavi, Juan Mata and Gerard Pique miss out, while Antonio Valencia is unavailable for Reinaldo Rueda's side
Del Bosque's side lost their last match, the Confederations Cup final against Brazil, ending a run of 23 games unbeaten and his fresh-faced squad will no doubt be designed to shake up the competition in the side.
Winger Antonio Valencia is the notable absentee from Reinaldo Rueda's Ecuador squad for the fixture in Guayaquil.
The Manchester United winger returned to domestic action in his side's 3-1 friendly loss to Sevilla before appearing in the 2-0 Community Shield win over Wigan, but Rueda believes using Valencia would be too much of a risk.
"Since June 11 when we played Argentina, Antonio hasn't played a single [international] match and that is a big difficulty," he told La Deportiva. "It is a risk for selection and for the player, because he is exposed to injury."
Ecuador have not won any of their last three matches, but remain in third position in the South America World Cup qualifying group with four games remaining.
The last time the two teams met was over a decade ago, with Spain coasting to a 4-0 victory thanks to a hat-trick from Fernando Morientes.
The fixture is Ecuador's first since the death of Christian Benitez earlier this month.