Although laparoendoscopic single-site surgery (LESS) is feasible among expert laparoscopic surgeons, it poses many technical challenges not seen in conventional laparoscopy (CL). Recent technological advancements in single-incision instrumentation have created more interest and widespread usage of LESS. However, neither LESS nor its novel instrumentation have been thoroughly studied or evaluated using human factors and ergonomics techniques. Consequently, the aim of this study was to compare the physical performance of LESS to CL using a standardized task. Wrist and elbow angular movements, range of motion and physical discomfort were assessed for 24 novice participants. There were no significant differences for physical comfort/discomfort ratings or elbow and wrist flexion/extension range of motion between CL and LESS. However, wrist radial/ulnar range of motion was significantly greater in LESS compared to CL (p < 0.05). Additionally, wrist radial/ ulnar range of motion was significantly greater using the SILS Port compared to the GelPOINT (p < 0.05). Although further investigation is needed, LESS resulted in greater wrist deviation and range of motion due to the close proximity of the instruments, restrictive nature of the single-port devices, and the need to achieve adequate instrument triangulation and visualization.