England defender Ashley Cole has been ruled out of Tuesday's World Cup qualifier against Ukraine, whilst John Terry faces a race to be fit.
Cole, 31, is suffering from an ankle injury, which forced him to sit out Friday's 5-0 win over Moldova.
The left-back was called-up to the original squad by Roy Hodgson, but returned to his club, Chelsea, for physiotherapy in the hope he could recover in time for the Ukraine match.
Meanwhile, centre-back Terry is receiving treatment for the an ankle injury he picked up against Moldova, and is struggling to be fit for England's second World Cup qualifier.
An FA statement read: "John Terry is continuing to receive treatment from the England medical team, having hurt his ankle in the World Cup qualifier with Moldova on Friday.
"Ashley Cole was assessed by the medical staff on Saturday and will not be available for the qualifier with Ukraine on Tuesday.''
Meanwhile, England midfielder Frank Lampard is looking forward to their tough encounter with Ukraine at Wembley.
Lampard, 34, netted twice in his country's 5-0 win over Moldova on Friday, as Roy Hodgson's men impressed in their opening World Cup qualifying.
The two teams met at Euro 2012 in a game that the Three Lions won 1-0 in Donetsk to see them through to the quarter finals.
The Chelsea man missed the major tournament during the summer through injury, and is hoping to make up for his absence at Wembley.
"Ukraine are a quality team, they showed that in glimpses in the game during the European Championship," Lampard told reporters. "It was a pretty impressive performance and a tough match for the team even though we got the win.
"They are certainly going to come to Wembley with intentions to make it difficult for us and show what they are about, so we will have to be on top form. We need to rest up, prepare right and try and keep the level of performance we are producing."
Gerrard, who put a fine performance alongside Lampard, said: "It will be a different test on Tuesday. With all due respect to Moldova, Ukraine are a fantastic team as they showed in the Euros. I'm sure it will be a lot tougher test."