| Place | Performance | Athlete | Team |
| 1 | 43.95 | Q: TEN CATE - AMY | WESTERN CAPE MAINSTREAM |
| 2 | 45.16 | Q: PIENAAR - DANYELLE | FREE STATE MAINSTREAM |
| 3 | 45.20 | Q: RAATH - CASSANDRA | GAUTENG MAINSTREAM |
| 4 | 45.25 | q: JENKINS - CAYLEN | GAUTENG MAINSTREAM |
| 5 | 45.34 | q: VERSTER - LEA | NORTH WEST MAINSTREAM |
| 6 | 45.42 | -: SMOOK - ZELMARIE JENNY | FREE STATE MAINSTREAM |
| 7 | 45.69 | -: LAMBRECHT - IZAAN | EASTERN CAPE MAINSTREAM |
| 8 | 48.53 | -: BOTES - KYLIE | LIMPOPO MAINSTREAM |