ASSAY OF THE ANTIANGIOGENIC COMPOUND TNP-470, AND ONE OF ITS METABOLITES, AGM-1883, BY REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY IN PLASMA

This paper describes a reversed-phase, high-performance liquid chromat ographic (HPLC) method for the isolation, detection, and quantificatio n of TNP-470 (I) and one of its active metabolites, AGM-1883 (II), fro m plasma. These compounds are initially extracted from plasma with an organic solvent and then separated from one another on a C18 column. T hose fractions eluting from the C18 column and containing either I or II are then derivatized through their epoxide moieties with sodium 8-q uinolinethiolate (SQT). This derivatization produces fluorescent speci es that are isolated and quantified by a second reversed-phase HPLC an alysis. The assay yields a lower limit of reliable quantification of 2 .5 ng/ml and is linear to a concentration at least as high as 160 ng/m l. The inter-assay percent coefficient of variation is less than 18%.