| 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 |