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To examine the associations of prolonged sedentary time (ST) with disease severity in women with fibromyalgia, and to analyse the combined association of total ST and prolonged ST with the disease severity in this population. Women (n = 451; 51.3 ± 7.6 years old) with fibromyalgia participated. Sedentary time and moderate-to-vigorous physical activity (MVPA) were measured using triaxial accelerometry and ST was processed into 30- and 60-min bouts. Dimensions of fibromyalgia (function, overall, symptoms) and the overall disease impact were assessed with the Revised Fibromyalgia Impact Questionnaire (FIQR). Body fat percentage was assessed using a bio-impedance analyser, and physical fitness was assessed with the Senior Fitness Tests Battery. Greater percentage of ST in 30-min bouts and 60-min bouts were associated with worse function, overall, symptoms and the overall impact of the disease (all, P < 0.05). Overall, these associations were statistically significant when additionally controlling for MVPA and overall physical fitness. Participants with low levels of total ST and prolonged ST (>60-min bouts) presented lower overall impact compared to participants with high levels of total ST and prolonged ST (mean difference = 6.56; 95% confidence interval (CI) = 1.83 to 11.29, P = 0.002). Greater percentage of ST accumulated in 30- and 60-min bouts and a combination of high levels of total and prolonged ST are related to worse disease severity. Although unable to conclude on causality, results suggest it might be advisable to motivate women with fibromyalgia to break prolonged ST and reduce their total daily ST. [ABSTRACT FROM AUTHOR]