Britain's Billy Harris and Anna Brogan missed out on places in the Wimbledon main draw by losing in the final qualifying round at Roehampton.
Harris, 28, lost 6-4 7-6 (8-6) 6-4 against big-serving Hamad Medjedovic from Serbia, before Brogan, 25, was beaten 6-3 6-0 by China's Bai Zhuoxuan.
It means there will be eight Britons in the men's singles event with six in the women's singles competition.
The main Wimbledon tournament begins on 3 July - with full coverage on the BBC.
Nineteen-year-old Medjedovic, ranked 155th in the world, got a break of serve in only the third game of the match and took the opening set, despite needing physio treatment on a right shoulder injury.
The Serb clinched the second set via a tie-break, then, following another lengthy medical timeout to deal with blisters on his feet, sealed the victory against Harris, ranked 345th, in three sets.
"It means the world to me - I still can't believe I've done it in my first time here," said Medjedovic.
"The serving was going quite good over the three rounds and I felt confident when serving. I had a very tough match yesterday - we played three hours, so not much time to recover and I'm still feeling pain in my body."
Qualifying took place at the Community Sports Centre in Roehampton and Brogan, 370th in the world and looking to qualify for her first Grand Slam, was unable to get past her 20-year opponent, who is ranked 191st.
"For me it's really amazing and why I started playing tennis - my dream is a Grand Slam," said Bai.
"She [Brogan] played good and we have known each other for a long time and are good friends. I just gave my best on court and was happy to be in control of the match."
Sixteen British players - eight men and eight women - had started qualifying, but none of them made it through the three rounds.
Britain's Dan Evans, Andy Murray and Cameron Norrie qualified automatically for the main draw, while Liam Broady, Jan Choinski, Arthur Fery, George Loffhagen and Ryan Peniston have been given wildcards.
No British women were ranked in the top 100 at the cut-off date for Wimbledon to qualify automatically, although Katie Boulter, Jodie Burrage, Harriet Dart, Sonay Kartal, Katie Swan and Heather Watson were handed wildcards.