Simultaneous determination of ITF 296 and two metabolites in plasma by a validated HPLC method.

A simple and sensitive HPLC validated method was developed for the simultaneous determination of ITF 296 (a new orally active nitrate) and its metabolites ITF 1124 and ITF 1577 in biological samples. Quantitation was performed by peak height ratios between the three compounds and the internal standard (ITF 1721). Detection limit of the method was 0.005 microgram/ml using 1 ml of plasma. The results indicated that the method is reproducible, accurate, precise, sensitive and specific for the measurement of the three compounds in plasma samples. Those characteristics allowed us to conclude that the method is suitable for analysing samples obtained after administration of ITF 296 to humans at therapeutic dosage and to animals for formulation studies.