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Date: 11 August 2008
Summary
Semi-final 1
| Rank | Name | Nation | Time |
| 1 | Arkady Vyatchanin | RUS | 53.06 |
| 2 | Aschwin Wildeboer | ESP | 53.51 |
| 3 | Liam Tancock | GBR | 53.61 |
| 4 | Junichi Miyashita | JPN | 53.69 |
| 5 | Tomomi Morita | JPN | 53.95 |
| 6 | Gregor Tait | GBR | 54.37 |
| 7 | Lubos Krizko | SVK | 54.38 |
| 8 | Mirco di Tora | ITA | 54.92 |
| Rank | Name | Nation | Time |
| 1 | Hayden Stoeckel | AUS | 52.97 |
| 2 | Matt Grevers | USA | 52.99 |
| 3 | Aaron Peirsol | USA | 53.56 |
| 4 | Ashley Delaney | AUS | 53.76 |
| 5 | Markus Rogan | AUT | 53.80 |
| 6 | Gerhard Zandberg | RSA | 53.98 |
| 7 | Stanislav Donets | RUS | 54.57 |
| 8 | Guy Barnea | ISR | 54.93 |