Natural organic matter (NOM) was isolated from the water of the Neversink Reservoir, part of the New York City water supply, located in the Catskill Mountains of New York. The NOM was fractionated into the following nine different fractions by the isolation procedure: (1) coarse particulates, (2) fine-particulate organics, (3) solvent-extractable organics, (4) hydrophobic neutrals (HPON fraction), (5) dissolved colloids, (6) bases, (7) hydrophobic acids (HPOA), (8) transphilic acids + neutrals (TPI-A+N), and (9) hydrophilic acids + neutrals (HPI-A+N). Each of these fractions, with exception of the first and the third which were too small for the complete series of analyses, was characterized by elemental, carbohydrate, and amino acid analyses, and by nuclear magnetic resonance and infrared spectrometry. The data obtained from these analyses indicate (1) that the fine-particulate organics and colloids are mainly composed of peptidoglycans, and lipopolysaccharides derived from algal, bacterial, and fungal cell walls, (2) that the HPO-N fraction most likely consists of a mixture of alicyclic terpenes and carbohydrates, (3) that the HPOA fraction consists mainly of lignin components conjugated to carbohydrates, (4) that the TPI-A+N and the HPI-A+N fractions most likely represent complex mixtures of relatively low molecular weight carboxylic acids derived from terpenes, carbohydrates, and peptides, and (5) that the base fraction is composed of free amino acids, browning reaction products, and peptide fragments.