Organic polyelectrolytes are utilized extensively in wastewater treatment, but their fate after use is poorly understood. Analytical methods used for polymer determination in less complex systems appear to fail in application to wastewater systems, contributing to the lack of knowledge. Thus, the development of 1H NMR spectroscopy is reported here for specifically quantitating certain cationic flocculant polymers in environmental samples. Proton observe frequencies of 250 or 400 MHz proton were used. A copolymer of acrylamide and acryloyloxyethyltrimethylammonium chloride was used, representative of cationic flocculant polymers possessing quaternary ammonium groups with terminal methyls that provide a sharp singlet at a chemical shift of approximately 3.06 ppm. A strong linear relationship was demonstrated between polymer concentration and either height or area of this peak. Recoveries were up to 96% at higher concentrations (250 mg/L), and were greater than when using viscosity or charge titration methods for polymer determination. Lesser recoveries at lower concentrations (70% at 5 mg/L) were attributed to adsorptive losses. The detection limit of this method was determined to be <0.5 mg/L. Use of the method was exemplified by analysis of anaerobically digested sludges for residual polymer following a range of dosages, showing the resulting isotherm.