Management of aquatic ecosystems is hampered because current methodology limits characterization of phosphorus (P) forms. We developed a procedure to characterize dissolved (DP) and particulate (PP) P from river waters by solution P-31 nuclear magnetic resonance (NMR) spectroscopy, using 4-L samples, and tested this procedure with a spiking trial. Most P was orthophosphate. Organic P forms included phosphonates, myo-inositol hexakisphosphate, and orthophosphate diesters. This research represents an important technical advance to characterize DP and PP in natural waters. It is simple, uses samples small enough for routine collection, and puts PP and DP into the same chemical environment for direct comparison. The technique is sensitive, detecting changes in spectra from P additions as small as 2% of total P, and identifying differences from two points along the flow path of a single river. However, lyophilizing samples in NaOH-ethylenediaminetetraacetic acid (EDTA) may alter some P forms, which requires further investigation.