Soil organic matter (OM) contains vast stores of carbon, and directly supports microbial, plant, and animal life by retaining essential nutrients and water in the soil. Soil OM plays important roles in biological, chemical, and physical processes within the soil, and arguably plays a major role in maintaining long-term ecological stability in a changing world. Despite its importance, there is a great deal still unknown about soil OM chemical ecology. The development of sophisticated analytical methods have reshaped our understanding of soil OM composition, which is now believed to be comprised of plant and microbial products at various stages of decomposition. The methods also have recently been applied to study environmental change in various settings and have provided unique insight with respect to soil OM chemical ecology. The goal of this review is to highlight the methods used to characterize soil OM structure, source, and degradation that have enabled precise observations of OM and associated ecological shifts. Although the chemistry of soil OM is important in its overall fate in ecosystems, the studies conducted to date suggest that ecological function is not defined by soil OM chemistry alone. The long-standing questions regarding soil OM stability and recalcitrance will likely be answered when several molecular methods are used in tandem to closely examine structure, source, age, degradation stage, and interactions of specific OM components in soil.