High speed HPLC determination of bis(pivaloyloxymethyl)-PMEA and its degradation products, mono(POM)-PMEA and PMEA

A high-speed HPLC gradient method for the determination of the antiviral ester prodrug bis(POM)-PMEA and its degradation products mono(POM)-PMEA and PMEA, is described. Perfusion chromatography enabled us to increase the flow rate up to 4 mL/min, resulting in high sample turn-over. Using a gradient system, the three compounds were separated from each other and the sample matrix within six minutes, while column equilibration was obtained in less than 3 minutes. Tetrabutylammonium was used as a counterion for PMEA. Notwithstanding the high flow rate, excellent reproducibility was obtained: the intraday as well as the interday precision, expressed as the relative standard deviation, for concentrations ranging from 5 to 50 mu M PMEA and bis(POM)-PMEA, was lower than 5%. The applicability of the method is demonstrated by studying the temperature- and pH-dependent degradation of bis(POM)-PMEA.