England head coach Simon Middleton has dropped three players for Saturday's Test against the United States at Exeter's Sandy Park.
Worcester prop Laura Keates, Gloucester full-back Emma Sing and Loughborough Lightning prop Detysha Harper miss out from the 35-woman squad.
The Test is part of preparations for the World Cup which begins in New Zealand on 8 October.
Middleton said those not selected remain "in contention".
"We've had a good month of preparation at Pennyhill Park, where the players have worked extremely hard," said Middleton.
"We've reduced the squad to 35 ahead of this weekend's match.
"We feel it's best for them to return to their clubs for their own pre-seasons, and we will of course be monitoring and in touch throughout."
England rose to the top of the world rankings in April with a fourth successive Six Nations title.
The Red Roses have a second warm-up game against Wales on 14 September in Bristol.
Middleton will then announce his final squad of 32 before they head out to New Zealand on Tuesday, 20 September.
England squad
Forwards: Zoe Aldcroft (Gloucester-Hartpury), Sarah Beckett (Gloucester-Hartpury), Sarah Bern (Bristol Bears), Hannah Botterman (Saracens), Shaunagh Brown (Harlequins), Bryony Cleall (Wasps), Poppy Cleall (Saracens), Amy Cokayne (Harlequins), Vickii Cornborough (Harlequins), Lark Davies (Bristol Bears), Vicky Fleetwood (Saracens), Rosie Galligan (Harlequins), Sarah Hunter (Loughborough Lightning), Sadia Kabeya (Loughborough Lightning), Alex Matthews (Gloucester-Hartpury), Maud Muir (Gloucester-Hartpury), Cath O'Donnell (Loughborough Lightning), Marlie Packer (Saracens), Connie Powell (Gloucester-Hartpury), Morwenna Talling (Loughborough Lightning), Abbie Ward (Bristol Bears)
Backs: Holly Aitchison (Saracens), Jess Breach (Saracens), Zoe Harrison (Saracens), Tatyana Heard (Gloucester-Hartpury), Natasha Hunt (Gloucester-Hartpury), Leanne Infante (Saracens), Ellie Kildunne (Harlequins), Claudia MacDonald (Exeter Chiefs), Sarah McKenna (Saracens), Lucy Packer (Harlequins), Amber Reed (Bristol Bears), Helena Rowland (Loughborough Lightning), Emily Scarratt (Loughborough Lightning), Lydia Thompson (University of Worcester Warriors)
Rehabilitation: Abby Dow (Wasps)