Quantitative assessment of tissue microstructure is important in studying human brain diseases and disorders in which white matter is implicated, as it has been linked to demyelination, re-myelination, and axonal damage in clinical conditions. Ultra-high field magnetic resonance imaging data obtained using a multi-echo gradient echo sequence has been shown to contain information on myelin, axonal and extracellular compartments in white matter. In this study, we aimed to assess the sensitivity of a three-compartment model to estimate the variation of corresponding compartment parameters (water fraction, relaxation time and frequency shift) of the corpus callosum sub-regions, which are known to have different tissue structure. Additionally, we computed the g-ratio using myelin and axonal water fractions and performed a voxel-by-voxel analysis in the corpus callosum. Based on data acquired for ten participants, we show that the myelin compartment water fraction and T2∗ is consistent across the corpus callosum sub-regions, whilst myelin frequency shift varies. The results show that the variation in water fraction, T2∗ and frequency shift for the myelin signal compartment across the corpus callosum is smaller than for the axonal and extracellular signal compartments. The computed g-ratio was comparable to previously published studies in the corpus callosum. Our study suggests that a multi-echo GRE approach in vivo combined with a complex three-compartment model is sensitive to microstructural parameter variations across the human corpus callosum.