Fatigue is a critical issue for female exercisers that may lead to medical errors, degradation in performance, decreased mental acuity, and social problems. Poor sleep quality is also a contributing factor to the fatigue experienced by female exercisers. Overwork is a factor causing poor sleep quality in female exercisers. The present study aimed to investigate the relationship between workload, fatigue, and sleep quality among female exercisers in the Sleman Regency District. A cross-sectional study was conducted from December 2023 to March 2024 using a self-reported questionnaire. The questionnaire included the Workload questionnaire adapted from the National Aeronautics and Space Administration Task Load Index, the Leeds Sleep Evaluation Questionnaire (LSEQ), and the Subjective Self Rating of Fatigue Assessment Scale (FAS) from the WHO Quality of Life Assessment Instrument (WHOQOL-EF) with 200 respondents in Sleman Regency. The PLS equation model was used to analyze the relationship between workload, sleep quality, fatigue, and the indicators of each instrument. The results showed that female exercisers who were over 40 years old, had children at home, had informal work, did daily exercise in the evening, and had a work schedule in the night had a higher risk of fatigue and poorer sleep quality than those below 40 years old who didn’t have children at home, had formal work, did daily exercise in the morning, and had a work schedule in the morning. A subjective self-rating test was used to measure fatigue, sleep quality, and workload. The training load and intensity of exercise programs influenced fatigue levels, which affected sleep quality. Female exercisers may face challenges balancing exercise with other commitments, such as family responsibilities, leading to overtraining and fatigue. The PLS model test showed a significant relationship between sleep quality and the level of perceived fatigue (p-value = 0.000). Workload significantly impacts sleep quality and fatigue risk in female exercisers. Therefore, managing workload is crucial to improving sleep quality and reducing fatigue risk.