A new approach to the qualitative and quantitative determination of nucleic acids is presented. Particle beam hollow cathode optical emission spectroscopy (PB/HC-OES) has been applied to determinations of five nucleotides (AMP, ADP, dATP, dCTP, and dGTP) by monitoring P I 219.9 nm and C I 193.0 nm optical emission. A high level of correlation exists between the phosphorus emission response and the degree of phosphorylation, demonstrating the technique’s ability to produce vital qualitative and quantitative information for specific nucleotides, with detection limits on the single-nanogram level for phosphorus (9 ng) and picogram level for carbon (20 pg), respectively, with RSD <12% for triplicate injections over a concentration range of 10−4–101 μg mL−1. The stoichiometric response suggests that it is possible to identify the various nucleotides by the amount of phosphate present within the nucleotide. Empirical formula calculations on P I and C I emission response ratios demonstrate the potential of the technique as an element specific detector for liquid chromatography (LC).