Igor Andreev, Svetlana Kuznetsova and Jelena Jankovic all bowed out of the AEGON International in the first round on Tuesday.
Andreev, the No.1 seed and world No.25, lost to Canadian qualifier Frank Dancevic 6-7(6) 2-6 in the opening match on Centre Court.
Dancevic, who is currently ranked No.126 in the world but has been as high as No.65, possesses an impressive grass court pedigree.
Last year, the 24 year-old won the $50k title in Surbiton before going on to defeat David Nalbandian in straight sets in the first round at Wimbledon.
Dancevic will face James Ward in the second round, the British No.4 who gained a wild card into the main draw and defeated top 100 player Victor Crivoi 6-1 6-3 in his opening match.
No.2 seed Kuznetsova, who is playing in her first grass court event since winning the French Open title, comfortably lost to another Canadian, Aleksandra Wozniak, 0-6 3-6.
It was an impressive performance from the 21 year-old who will now meet China’s Jie Zheng in the second round – both players reached the third round at last week’s AEGON Classic in Edgbaston.
No.3 seed Jankovic, a former world No.1, won the opening set of her first round match but was eventually knocked out, losing to Russian Anna Chakvetadze 7-6(5) 3-6 2-6 in over two hours.
“I didn't really feel comfortable out there,” said Jankovic. “I got into the match at the start and won the first set and felt I was playing quite well. But the ball stayed so low and my legs felt very tired and I didn't want to bend down for the balls so I lost my rhythm.”
Chakvetadze now goes on to face Frenchwoman Marion Bartoli, the runner-up at Wimbledon in 2007, in a match the 22 year-old is looking forward to playing.
"Marion is a very tough opponent on grass and she has done well at Wimbledon in the past,” said Chakvetadze. “I'll just have to see how it goes and try to play my best tennis. I am looking forward to the match."
Elsewhere in the women’s draw, reigning champion Agnieszka Radwanska of Poland is safely through to the second round after a straight sets victory over her younger sister Urszula while Caroline Wozniacki and Amelie Mauresmo also progressed on Tuesday.
In the men’s event, three second round matches were completed with the No.4 seed from France, Fabrice Santoro, beating Robbie Ginepri 4-6 6-4 6-1, Guillermo Garcia-Lopez beating Yen-Hsun Lu 6-4 6-7(0) 7-6(3) and Ivan Ljubicic beating Josh Goodall 6-3 7-5.
“This week was my first tour event win,” said Goodall, the British No.3. “I’ve had about nine chances but I’ve had some difficult draws. It’s is a big confidence booster, especially after Nottingham where I made semi-finals a couple of weeks ago. I’ve prepared myself well for Wimbledon.”
On Monday evening, Anne Keothavong and Elena Baltacha are both getting their singles campaigns under way at Devonshire Park against Austria’s Sybille Bammer and China’s Na Li respectively.