Background: Objective sensor-based quantification of sedentary behavior is an important tool for planning and evaluating interventions for excessive sedentary behavior in patients with musculoskeletal diseases. Although waist-worn accelerometers are the standard for physical activity (PA) assessment, only thigh-worn inclinometers can clearly distinguish sedentary behavior from any light PA or standing activity. Methods: In this study, 53 adults (ages 20–85 years) wore two ActiGraph wGT3X-BT monitors, each containing an inclinometer and accelerometer (set for acquisition of slow movements in all three planes), attached to the right waist and thigh for a period of about 4 days. Both monitors recorded total sedentary time and continuous sedentary 10-min bouts by synchronous accelerometry and inclinometry. Differences and correlations between methods and wearing positions were evaluated against participant age, body mass index (BMI), and number of steps taken. Thigh-worn inclinometry was used as reference. Results: Data from thigh-worn inclinometry and waist-worn accelerometry were highly correlated for total sedentary time [rho = 0.888; intraclass correlation coefficient (ICC) = 0.937] and time in sedentary bouts (rho = 0.818; ICC = 0.848). Nevertheless, accelerometry at the waist underestimated sedentary time by ≈17% (p < 0.001) and time in sedentary bouts by ≈54% (p < 0.001). A satisfactory concordance thus could be demonstrated only for total sedentary time, based on the Bland–Altmann method (≈96% of data within the limits of agreement). The differences between waist-worn accelerometry and thigh-worn inclinometry did not correlate with age but did correlate with BMI and PA for both sedentary behavior parameters (r ⩾ 0.240, p ⩽ 0.043). Conclusion: A waist-worn accelerometer can be used to determine total sedentary time under free-living conditions with sufficient accuracy if the correct settings are chosen. Further investigations are necessary to investigate why short sedentary bouts cannot be reliably assessed.