Mise au point et optimisation d’une méthode de dosage du cyclophosphamide par hplc dans les préparations magistrales pédiatriques

Abstract

Cyclophosphamide is an alkylating agent used in cancer chemotherapy. It acts by inhibiting DNA replication in rapidly dividing cells. In order to develop an HPLC method for determining the retention time of cyclophosphamide, a three-stage final screening design study was carried out. The factors studied were the composition of the mobile phase (percentage of acetonitrile) and the flow rate. The responses analysed were the number of theoretical plats, retention time and resolution with adjacent peaks. The analysis showed that the composition of the mobile phase was the most influential factor on the number of theoretical plateaus, with optimum conditions at 37% acetonitrile. The flow rate and composition of the mobile phase were the most influential factors on retention time, with optimum conditions at 20% acetonitrile and 1 mL/min. For resolution, the flow rate and composition of the mobile phase, as well as their interaction, were significant, with optimum conditions at 34% acetonitrile and 1 mL/min. An MODR was used to define a robust design space, in which an experimental point was chosen (1.49 mL/min, 25% acetonitrile) to avoid any drift during the routine.