The soap couple, who play Debbie Dingle and David Metcalfe in the long-running ITV show, had a son called Buster in 2010.
Charley Webb and Matthew Wolfenden
A spokeswoman for the show said: "Charley Webb and Matthew Wolfenden have today confirmed that they have separated. They will remain friends and are focusing on their little boy."
The pair, who got engaged in 2009, are both expected to remain with the hit show which celebrated 40 years on air last year.
Matthew and Charley in happier times
Matthew, who won reality show 'Dancing On Ice' in 2012, tweeted on Wednesday:
Charley's brother, Jamie Lomas, is married to 'Coronation Street' star Kym Marsh and recently signed up to appear in 'EastEnders'.