Real Madrid forward Gareth Bale will miss Wales' friendly against Netherlands on Nov. 13, although reports suggest he is not a doubt for the Clasico with Barcelona on Nov. 21.
Bale, 26, has not played since Oct. 17 after suffering a recurrence of a calf injury following the international break.
The winger featured in the first half of Real's 3-0 victory over Levante four days after Wales' 2-0 victory over Andorra having also played as they sealed a place at Euro 2016 despite losing 2-0 against Bosnia & Herzegovina.
The Welshman was criticised by some Madrid journalists and fans for playing for his country so soon after being sidelined for nearly three weeks with a calf strain.
Arsenal manager Arsene Wenger also hit out at Wales boss Chris Coleman for playing Aaron Ramsey against Andorra with Wales having already qualified for next summer's European Championship.
Ramsey subsequently picked up a hamstring injury on Arsenal duty and also misses Wales' first Euro 2016 warmup game at the Cardiff City Stadium.
Reading forward Hal Robson-Kanu and Fulham defender Jazz Richards are other absentees through injury. Newcastle defender Paul Dummett and Walsall striker Tom Bradshaw have been called into the squad.
Wales squad: Hennessey (Crystal Palace), Fon Williams (Inverness), Ward (Liverpool); A Williams (Swansea, capt), Chester (West Brom), Gunter (Reading), Taylor (Swansea), Davies (Tottenham), Collins (West Ham), Henley (Blackburn), Dummett (Newcastle), Vaughan (Nottingham Forest), Huws (Wigan), Allen (Liverpool), Ledley (Crystal Palace), King (Leicester), Edwards (Wolves), J Williams (Crystal Palace), Cotterill (Birmingham), Lawrence (Leicester), Church (MK Dons), Vokes (Burnley), Bradshaw (Walsall).