No British women will feature in the main French Open draw - the first time that has occurred in a Grand Slam since the 2009 US Open.
Katie Boulter, Harriet Dart and Fran Jones were eliminated in the second qualifying round, following Heather Watson, Katie Swan and Lily Miyazaki in missing out on a spot in the main tournament next week.
Emma Raducanu is out of the event after undergoing ankle and wrist operations.
Boulter lost a three-setter to America's Ashlyn Krueger on Wednesday, beaten 6-2 2-6 7-6 (10-3), while Dart fell to France's Elsa Jacquemot 1-6 6-1 6-2 as she was unable to build on a comprehensive first set.
Mayar Sherif: The history-maker blazing a trail for Arab womenMarion Bartoli: Emma Raducanu should not rush backJohanna Konta: Emma Raducanu can come back strongerJones had beaten two-time Grand Slam semi-finalist Coco Vandeweghe on Monday but saw her hopes of reaching the main stage at Roland Garros ended when she retired hurt because of a shoulder injury against Switzerland's Ylena In-Albon.
Cameron Norrie, Dan Evans and Jack Draper will be the only British players to feature in singles at the second Grand Slam of the year.
Andy Murray opted out in order to prioritise the grass-court season and Kyle Edmund withdrew shortly before the draw was made, while Liam Broady, Jan Choinski and Ryan Peniston lost in qualifying.
The draw for the French Open takes place on Thursday afternoon with Carlos Alcaraz the top seed for the men's tournament and Iga Swiatek taking that spot in the women's.