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Date: 14 August 2008
Summary
Semi-final 1
| Rank | Name | Nation | Time |
| 1 | Natalie Coughlin | USA | 53.70 |
| 2 | Zhu Yingwen | CHN | 53.84 |
| 3 | Britta Steffen | GER | 53.96 |
| 4 | Lisbeth Trickett | AUS | 54.10 |
| 5 | Josefin Lillhage | SWE | 54.59 |
| 6 | Lacey Nymeyer | USA | 54.74 |
| 7 | Petra Dallmann | GER | 55.05 |
| 8 | Aleksandra Gerasimenya | BLR | 55.31 |
| Rank | Name | Nation | Time |
| 1 | Marleen Veldhuis | NED | 53.81 |
| 2 | Hanna-Maria Seppala | FIN | 53.84 |
| 3 | Francesca Halsall | GBR | 53.94 |
| 4 | Jeanette Ottesen | DEN | 54.05 |
| 5 | Malia Metella | FRA | 54.20 |
| 6 | Cate Campbell | AUS | 54.54 |
| 7 | Erica Morningstar | CAN | 55.36 |
| Disqualified | |||
| Pang Jiaying | CHN | ||