Phosphoramide mustard (PM), a key active metabolite of the widely used anticancer drug cyclophosphamide (CP), can exist in several closely-related ionized, cyclized and substituted forms. We have developed a high pressure liquid chromatographic (HPLC) method for analysing serum concentrations of PM in order to relate these serum concentrations to toxicity and efficacy of treatment of CP.31 p NMR spectroscopy is used to verify the HPLC peak homology of the proposed PM peak. © 1987, Taylor & Francis Group, LLC. All rights reserved.