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Date: 16 September 2000
Summary
Semi-final 1
| Rank | Name | Nation | Time |
| 1 | Jenny Thompson | USA | 58.18 |
| 2 | Martina Moravcova | SVK | 58.49 |
| 3 | Diana Mocanu | ROU | 59.12 |
| 4 | Otylia Jedrzejczak | POL | 59.14 |
| 5 | Natalia Soutiaguina | RUS | 59.30 |
| 6 | Mette Jacobsen | DEN | 59.75 |
| 7 | Cecile Jeanson | FRA | 59.80 |
| 8 | Anna-Karin Kammerling | SWE | 1:00.40 |
| Rank | Name | Nation | Time |
| 1 | Inge de Bruijn | NED | 57.14 |
| 2 | Petria Thomas | AUS | 58.11 |
| 3 | Dara Torres | USA | 58.35 |
| 4 | Junko Onishi | JPN | 59.04 |
| 5 | Susie O'Neill | AUS | 59.05 |
| 6 | Johanna Sjoeberg | SWE | 59.15 |
| 7 | Amanda Loots | RSA | 59.63 |
| 8 | Sophia Skou | DEN | 59.89 |