Striker Hal Robson Kanu has retired from international football with immediate effect.
Robson-Kanu and defender Neil Taylor were absentees from the Wales squad to play the Republic of Ireland and Denmark in Nations League qualifiers.
The 29-year old West Brom forward confirmed the news in a statement shortly after the squad announcement.
Newcastle defender Paul Dummett, who previously asked not to be selected for international football, is recalled.
Robson-Kanu retires having made 44 Wales appearances, scoring five goals, including two at the Euro 2016 finals. Most famously, he scored Wales' second goal in their stunning 3-1 win over Belgium in the quarter-finals of that tournament.
"I have made the decision to retire from international football. Long thought, but a decision made; commitments to my family and dedicating what time I have available to them and choosing to focus on my club career within sport and time needed away from the game," Robson-Kanu wrote.
"I wish Ryan Giggs and the team continued success in the years to come, and appreciate his understanding and support, a new generation to continue building on what Gary Speed and Chris Coleman have set in place."
Manager Giggs says he understands the decision made by the West Brom forward, who he believes has secured his status as a Welsh football "icon".
"I spoke to him yesterday and it was a difficult decision for him," Giggs said.
"He's had a difficult six months, with the birth of his baby, and he wants to focus on his family and his club career.
"He's an icon in Welsh football with the goal he scored against Belgium and I wish him all the best."
Teenage forward Tyler Roberts is selected in the Wales squad, as is Manchester City's 18-year old Matthew Smith and Chelsea defender Ethan Ampadu.
However, Euro 2016 squad member Jazz Richards misses out.
Wales manager Giggs also confirmed defender Ashley Williams will remain as Wales captain despite being sent off in two of his past four club appearances.
"Ash has been fantastic since I took over, leader on and off the pitch," said Giggs.
"Nothing's changed in that respect and it's good to see him playing regular now at Stoke."
Giggs said he was pleased to be able to recall Dummett, who asked not to be selected for a qualifier in Serbia in 2017.
"It is a boost. I spoke to Paul when I took over and he wanted to focus on Newcastle at the time.
"But I've had a chat with Paul and he's in a good place now.
"He's playing regularly and captained his boyhood club on the weekend. I'm looking forward to working with him."
Wales squad: Hennessey, Ward, Adam Davies, Gunter, Roberts, Chester, Williams, Mepham, Lockyer, Ben Davies, Dummett, John, Ampadu, Allen, Smith, Ramsey, King, Ledley, Wilson, Brooks, Lawrence, Woodburn, Bale, Vokes, Roberts.