Polymeric prodrugs (polymer-drug conjugates) are unique and can provide various advantagesover other nanotechnology platforms. Rather than encapsulation of drug in a nanoparticle shell, polymeric prodrugs use covalent conjugation to bind the drug, or active pharmaceutical ingredient (API), to the polymer backbones. Additionally, the drug loading ratio and subsequent drug release rates can be optimized by tailoring the chemistry used to incorporate the APIs. Unlike the encapsulation of APIs in the other nano-sized drug delivery systems, the quantification of chemically conjugated drugs in polymeric prodrugs is more difficult. Here, we describe a protocol for drug quantification in polymeric prodrugs using an automated elemental analyzer (PE 2400 Series II CHNS/O, Perkin Elmer). Briefly, materials containing carbon (C), hydrogen (H), nitrogen (N), and/or sulfur (S) are combusted under high temperatureand high oxygen conditions into their oxidized gas form, then analyzed by internal gas chromatography (GC) for a quantitative analysis which is reported as a percentage of the original sample weight. This document summarizes the laboratory procedure. For a more descriptive review of the procedure, we refer you to the publication “Total Drug Quantification in Prodrugs Using an Automated Elemental Analyzer” which describes the methods and related analysis to measure the amount of drug loading in a polymeric drug conjugate using poly(L-lysine succinylated) (PLS) and lamivudine (LAM).