| Ranking | time | gap | React | HR |
| 1. Pernille Blume | DEN | 24.23s | 0.63 | Q | 1. |
| 2. Fran Halsall | GBR | 24.26s | 0.03 | 0.70 | 1. |
| 3. Aliaksandra Herasimenia | BLR | 24.42s | 0.19 | 0.70 | 1. |
| 4. Bronte Campbell | AUS | 24.45s | 0.22 | 0.67 | 2. |
| 5. Jeanette Ottesen | DEN | 24.48s | 0.25 | 0.64 | 2. |
| 5. Abbey Weitzeil | USA | 24.48s | 0.25 | 0.67 | 2. |
| 7. Cate Campbell | AUS | 24.52s | 0.29 | 0.82 | 3. |
| 8. Ranomi Kromowidjojo | NED | 24.57s | 0.34 | 0.67 | 3. |
| 8. Chantal Jean van Landeghem | CAN | 24.57s | 0.34 | 0.69 | 3. |
| 10.Sarah Sjostrom | SWE | 24.66s | 0.43 | 0.70 | 3. |
| 11.Simone Manuel | USA | 24.71s | 0.48 | 0.69 | 4. |
| 12.Therese Alshammar | SWE | 24.73s | 0.50 | 0.62 | 5. |
| 13.Dorothea Brandt | GER | 24.77s | 0.54 | 0.67 | 4. |
| 13.Inge Dekker | NED | 24.77s | 0.54 | 0.69 | 1. |
| 13.Arianna Vanderpool-Wallace | BAH | 24.77s | 0.54 | 0.62 | 4. |
| 16.Etiene Medeiros | BRA | 24.82s | 0.59 | 0.61 | 6. |
| eliminated |
| 17.Silvia di Pietro | ITA | 24.89s | 0.66 | 0.64 | 7. |
| 18.Xiang Liu | CHN | 24.91s | 0.68 | 0.68 | 5. |
| 18.Farida Hisham Ahmed Osman | EGY | 24.91s | 0.68 | 0.70 | 2. |
| 18.Michelle Williams | CAN | 24.91s | 0.68 | 0.70 | 6. |
| 21.Anna Santamans | FRA | 24.93s | 0.70 | 0.60 | 6. |
| 22.Vanessa Garcia Vega | PUR | 24.94s | 0.71 | 0.63 | 1. |
| 22.Rozaliya Nasretdinova | RUS | 24.94s | 0.71 | 0.70 | 8. |
| 24.Susann Bjornsen | NOR | 25.05s | 0.82 | 0.72 | 1. |
| 25.Flora Molnar | HUN | 25.07s | 0.84 | 0.65 | 7. |
| 26.Julie Meynen | LUX | 25.12s | 0.89 | 0.62 | 2. |
| 27.Yuliya Khitraya | BLR | 25.18s | 0.95 | 0.70 | 3. |
| 28.Liliana Ibanez Lopez | MEX | 25.25s | 1.02 | 0.71 | 3. |
| bellow Olympic Qualifying Time - 25.28s |
| 29.Aleksandra Urbanczyk | POL | 25.28s | 1.05 | 0.66 | 4. |
| 30.Isabella Arcila Hurtado | COL | 25.35s | 1.12 | 0.61 | 4. |
| 31.Theodora Drakou | GRE | 25.36s | 1.13 | 0.62 | 5. |
| 31.Melanie Henique | FRA | 25.36s | 1.13 | 0.62 | 2. |
| 33.Zohar Shikler | ISR | 25.38s | 1.15 | 0.79 | 3. |
| 34.Erika Ferraioli | ITA | 25.40s | 1.17 | 0.69 | 4. |
| 35.Andrea Murez | ISR | 25.41s | 1.18 | 0.75 | 6. |
| 36.Rikako Ikee | JPN | 25.45s | 1.22 | 0.66 | 7. |
| 37.Anna Dowgiert | POL | 25.54s | 1.31 | 0.65 | 5. |
| 38.Natalia Lovtcova | RUS | 25.55s | 1.32 | 0.76 | 7. |
| 39.Birgit Koschischek | AUT | 25.58s | 1.35 | 0.68 | 6. |
| 40.Graciele Herrmann | BRA | 25.60s | 1.37 | 0.65 | 8. |
| 41.Alexandra Sasha Touretski | SUI | 25.66s | 1.43 | 0.63 | 5. |
| 42.Darya Stepanyuk | UKR | 25.67s | 1.44 | 0.73 | 8. |
| 43.Yayoi Matsumoto | JPN | 25.73s | 1.50 | 0.73 | 7. |
| 44.Camille Lily Mei Cheng | HKG | 25.92s | 1.69 | 0.66 | 6. |
| 45.Allyson Ponson | ARU | 26.00s | 1.77 | 0.72 | 7. |
| 46.Karen Torrez | BOL | 26.16s | 1.93 | 0.62 | 1. |
| bellow Olympic Selection Time - 26.17s |
| 47.Naomi Ruele | BOT | 26.23s | 2.00 | 0.70 | 2. |
| 48.Elinah Phillip | IVB | 26.26s | 2.03 | 0.63 | 3. |
| 49.Bayan Jumah | SYR | 26.41s | 2.18 | 0.71 | 4. |
| 49.Pei Wun LIN | TPE | 26.41s | 2.18 | 0.74 | 8. |
| 51.Talita Baqlah | JOR | 26.48s | 2.25 | 0.78 | 5. |
| 52.Bexx Heyliger | BER | 26.54s | 2.31 | 0.68 | 6. |
| 53.Nikki Muscat | MLT | 26.60s | 2.37 | 0.71 | 7. |
| 54.Faye Hussain | KUW | 26.86s | 2.63 | 0.73 | 1. |
| 55.Dorian McMENEMY | DOM | 27.37s | 3.14 | 0.74 | 8. |
| 56.Naomy Hope Grand-Pierre | HAI | 27.46s | 3.23 | 0.73 | 2. |
| 57.Samantha Roberts | ANT | 27.95s | 3.72 | 0.83 | 3. |
| 58.Colleen Furgeson | MHL | 28.16s | 3.93 | 0.72 | 1. |
| 59.Noura ???? | MAR | 28.20s | 3.97 | 0.69 | 4. |
| 60.Yesui Bayar | MGL | 28.40s | 4.17 | 0.74 | 2. |
| 61.Irene Prescott | TGA | 28.68s | 4.45 | 0.81 | 5. |
| 62.Miri Alatrash | PLE | 28.76s | 4.53 | 0.73 | 3. |
| 63.Jamila Sanmoogan | GUY | 28.88s | 4.65 | 0.72 | 6. |
| 64.Lianna Catherine Swan | PAK | 29.02s | 4.79 | 0.66 | 7. |
| 65.Dirngulbai Misech | PLW | 29.19s | 4.96 | 0.71 | 1. |
| 66.Vitiny Hemthon | CAM | 29.37s | 5.14 | 0.70 | 4. |
| 67.Magdalena Moshi | TAN | 29.44s | 5.21 | 0.80 | 5. |
| 67.Angelika Ouedraogo | BUR | 29.44s | 5.21 | 0.80 | 2. |
| 69.Sonia Aktar | BAN | 29.99s | 5.76 | 0.82 | 3. |
| 70.Thet Ei Ei | MYA | 30.25s | 6.02 | 0.88 | 1. |
| 71.Ammara Pinto | MAW | 30.32s | 6.09 | 0.64 | 6. |
| 72.Debra Daniel | FSM | 30.83s | 6.60 | 0.75 | 7. |
| 73.Anastasiya Tyurina | TJK | 31.15s | 6.92 | 0.78 | 1. |
| 74.Fatema Abdulmohsen Ahmed Almahmeed | BRN | 32.28s | 8.05 | 0.76 | 2. |
| 75.Rahel Fseha Gebresilassie | ETH | 32.51s | 8.28 | 0.91 | 4. |
| 76.Siri Arun Budcharern | LAO | 32.55s | 8.32 | 0.75 | 5. |
| 77.Laraiba Seibou | BEN | 33.01s | 8.78 | 0.82 | 2. |
| 78.Nada Mohamed Abdulzaher Mohamed Albedwawi | UAE | 33.42s | 9.19 | 0.70 | 3. |
| 79.Rebecca Kpossi | TOG | 33.44s | 9.21 | 0.83 | 6. |
| 80.Elsie Uwamahoro | BDI | 33.70s | 9.47 | 0.83 | 7. |
| 81.Fatoumata Samassekou | MLI | 33.71s | 9.48 | 0.89 | 8. |
| 81.Stefan Sangala | CGO | 33.71s | 9.48 | 0.75 | 3. |
| 83.Roukaya Moussa Mahamane | NIG | 35.60s | 11.37 | 0.89 | 4. |
| 84.Haneen Ibrahim | SUD | 36.23s | 12.00 | 0.86 | 5. |
| 85.Chloe Sauvourel | CAF | 37.15s | 12.92 | 0.98 | 6. |
| 86.Mohamed Andhumdine Nazlati | COM | 37.66s | 13.43 | 0.73 | 7. |
| 87.Mariama Dioulde Sow | GUI | 39.85s | 15.62 | 0.85 | 8. |
| 88.Bunturabie Effuah Rashida Jalloh | SLE | 39.93s | 15.70 | 0.88 | 4. |
| -. Awa Ly Ndiaye | SEN | DSQ |
| -. Xinyi Chen | CHN | DNS |
| -. Monika Vasiliyan | ARM | DNS |