Background Many workers performing manual handling tasks suffer from musculoskeletal disorders (MSD). Previous research has identified several loading aspects associated with manual handling, but it is still unknown if lifting on an unstable surface is associated with increased biomechanical loading of different body parts. Aim This proof-of-concept study aims to study what kinematic and kinetic movement parameters, such as movement time, joint angles, torque, and muscle activity are feasible and of importance when studying the effect of lifting on surfaces with varying degrees of stability in an experimental set-up. Methods Measurements were taken during three different surface conditions: stable, slightly unstable, and unstable. The participants were instructed to lift a box from the floor and place it on a table in front of them. The weight of the box varied from 0.5 to 15.5 kg. By using a motion capture system (VICON) with 28 reflective markers placed on the participants and one on the box, one Kistler force plate for measuring force levels and center of pressure movements (CoP), and four electromyographic transmitters (EMG), we analyzed the downward and upward phases of the lifting movement, using the Friedman’s test for repeated measures. Results Statistically significant results with less joint movements in the lower and upper back were seen with increased instability during both the downward and upward phases. The decrease in trunk movements with increased instability resulted in a somewhat more flexed knee position during the movement, a lower torque in the lower back, and a decrease in CoP movements, but no differences in movement time or muscle activity in back and knee muscles. Conclusion Lifting while standing on unstable surfaces resulted in an alteration of both kinematics and kinetics parameters; however, further studies regarding whether this is an additional risk factor for developing lower back pain are needed. Muscle activity levels were not altered due to instability and due to the complexity of the measurement, and we suggest not including EMG measures in future experiments of this type.