The two midfielders earn their first call-ups to the senior side after impressing in the opening two games of the Premier League season, with Rickie Lambert again included
The Spurs winger scored twice against Dinamo Tbilisi on his first start for the club before winning the decisive penalty in the 1-0 win over Swansea City on Sunday.
Barkley, meanwhile, opened his account for Everton in the 2-2 draw with Norwich City on the opening day of the season, before impressing against West Brom at the weekend.
Elsewhere, Fraser Forster returns to the international set-up, in place of Ben Foster - who was ruled out of action for three months on Tuesday after suffering a stress fracture in his right foot.
Rickie Lambert, who scored on his England debut, is awarded with a second call-up by Roy Hodgson, and joins strikers Wayne Rooney, Jermain Defoe and Daniel Sturridge - who has two goals in as many Premier League games this season - in the squad.
England host Moldova at Wembley on September 6 before facing Ukraine in Kyiv on September 10.
Goalkeepers: Joe Hart, John Ruddy, Fraser Forster.
Defenders: Leighton Baines, Gary Cahill, Ashley Cole, Phil Jagielka, Glen Johnson, Phil Jones, Chris Smalling, Kyle Walker.
Midfielders: Ross Barkley, Michael Carrick, Tom Cleverley, Steven Gerrard, Frank Lampard, James Milner, Raheem Sterling, Andros Townsend, Theo Walcott, Jack Wilshere.
Forwards: Jermain Defoe, Rickie Lambert, Wayne Rooney, Daniel Sturridge, Danny Welbeck, Ashley Young.