Applying two-dimensional correlation spectroscopy to (13)C NMR and FTIR spectra of the high molecular-weight dissolved organic matter (HMW-DOM) isolated along an Elizabeth River/Chesapeake Bay salinity transect shows that HMW-DOM consists of three major components that have different biogeochemical reactivities. The first appears to be a heteropolysaccharide (HPS) component and its contribution to carbon increases as we approach the marine offshore. The second appears to be composed of carboxyl-rich compounds (CRC); its carbon percentage decreases. The third component contains the major functional group of amide/amino sugar (AMS) and its carbon percentage stays almost constant along the salinity transect. It seems that the HPS and CRC are present in many aquatic environments at different relative ratios. The 2D-correlation maps reveal that each of these components is composed of dynamic mixtures of compounds that share similar backbone structures but have significant functional group differences. Two-dimensional (2D) correlation spectroscopy is a powerful new biogeochemical tool to track the changes in complex organic matter as a function of space, time, or environmental effects.