| Place | Performance | Athlete | Team |
| 1 | 25.13 | Q: BOOYSEN - MIENKE | MPUMALANGA MAINSTREAM |
| 2 | 25.31 | Q: DE KOCK - ASHTON | GAUTENG MAINSTREAM |
| 3 | 25.57 | Q: KOTZE - DANE | GAUTENG MAINSTREAM |
| 4 | 25.61 | -: ENGELBRECHT - MONIQUE | WESTERN CAPE MAINSTREAM |
| 5 | 25.72 | -: GEORGE - ANNESTAYSHA | EASTERN CAPE MAINSTREAM |
| 6 | 26.00 | -: PRINSLOO - MEGAN | LIMPOPO MAINSTREAM |
| 7 | 27.76 | -: DEVENISH - LIANE | NORTHERN CAPE MAINSTREAM |