The neurobiology of handedness is still poorly understood in nonhuman primates. Recently, an association between hand preference and precentral gyrus morphology in chimpanzees was reported. The aim of this study was to further evaluate the association between handedness and asymmetries in the precentral gyrus of chimpanzees (Pan troglodytes) and to evaluate the association between hand preference and brain asymmetry using a different approach to the classification of handedness in chimpanzees. The overall results suggest that differences in handedness groups are specific to a region of the precentral gyrus commonly known as the "knob" and that subjects that show different hand preferences differ in brain asymmetries for specific regions of the primary motor cortex. Moreover, using a continuous scale of measurement rather than discrete classification of handedness, significant associations were found between hand use and asymmetries within the precentral gyrus.