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Date: 13 August 2008
Summary
Semi-final 1
| Rank | Name | Nation | Time |
| 1 | Alain Bernard | FRA | 47.20 |
| 2 | Stefan Nystrand | SWE | 47.91 |
| 3 | Jason Lezak | USA | 47.98 |
| 4 | Lyndon Ferns | RSA | 48.00 |
| 5 | Cesar Cielo Filho | BRA | 48.07 |
| 6 | Christian Galenda | ITA | 48.47 |
| 7 | Jonas Persson | SWE | 48.59 |
| 8 | Fabien Gilot | FRA | 49.00 |
| Rank | Name | Nation | Time |
| 1 | Eamon Sullivan | AUS | 47.05 |
| 2 | Pieter van den Hoogenband | NED | 47.68 |
| 3 | Matt Targett | AUS | 47.88 |
| 4 | Filippo Magnini | ITA | 48.11 |
| 5 | Garrett Weber-Gale | USA | 48.12 |
| 6 | Brent Hayden | CAN | 48.20 |
| 7 | Andrey Grechin | RUS | 48.71 |
| 8 | Dominik Meichtry | SUI | 49.58 |