A series of graphite nanofibers (GNFs) that were subjected to various pretreatments were used to determine how modifications in the carbon structure formed during either synthesis or pretreatment steps results in active or inactive materials for hydrogen storage. The nanofibers possessing a herringbone structure and a high degree of defects were found to exhibit the best performance for hydrogen storage. These materials were exposed to several pretreatment procedures, including oxidative, reductive, and inert environments. Significant hydrogen storage levels were found for several in situ pretreatments. Examination of the nanofibers by high-resolution transmission electron microscopy (TEM) after pretreatment and subsequent hydrogen storage revealed the existence of edge attack and an enhancement in the generation of structural defects. These findings suggest that pretreatment in certain environments results in the creation of catalytic sites that are favorable toward hydrogen storage. The best pretreatment resulted in a 3.8% hydrogen release after exposure at 69 bar and room temperature.