Sedentary Behavior (SB) is among the most frequent human behaviors and is associated with a plethora of serious chronic lifestyle diseases as well as premature death. Office workers in particular are at an increased risk due to their extensive amounts of occupational SB. However, we still lack an objective method to measure SB consistent with its definition. We have therefore developed a new measurement system based on muscular activity and accelerometry. The primary aim of the present study was to calibrate the new-developed 8-CH-EMG+ for measuring occupational SB against an indirect calorimeter during typical desk-based office work activities. In total, 25 volunteers performed nine office tasks at three typical workplaces. Minute-by-minute posture and activity classification was performed using subsequent decision trees developed with artificial intelligence data processing techniques. The 8-CH-EMG+ successfully identified all sitting episodes (AUC = 1.0). Furthermore, depending on the number of electromyography channels included, the device has a sensitivity of 83–98% and 74–98% to detect SB and active sitting (AUC = 0.85–0.91). The 8-CH-EMG+ advances the field of objective SB measurements by combining accelerometry with muscular activity. Future field studies should consider the use of EMG sensors to record SB in line with its definition. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.