Lee Dickson and Danny Care were named on Tuesday as the two scrumhalves in a 26-man squad and young wingers Jack Nowell, Jonny May and Anthony Watson were preferred to Ashton.
"Competition at scrumhalf has been particularly tight and Danny's form leading into the Six Nations has been excellent and he merits his inclusion," England coach Stuart Lancaster said.
"Ben misses out this time but he is a quality player and I am sure he will respond in the same way Danny did at the start of the year.
"We have been pleased with the form of Chris (Ashton) recently. However, Jack has made great progress on the right wing and has also delivered in big games for Exeter Chiefs and we feel it is the right time to look at another option in that position."
Brad Barritt, Billy Twelvetrees and Luther Burrell were the three centres included in another area of intense competition for places.
(Reporting by Ed Osmond, editing by Ken Ferris)
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