Sensitive High-performance Liquid Chromatographic Assay for Endralazine and Two of Its Metabolites in Human Plasma

Endralazine (I) is a new antihypertensive which is chemically and pharmacologically related to hydralazine and dihydralazine. A sensitive high-performance liquid chromatographic-fluorescence assay for the drug and two of its metabolites [methyltriazoloendralazine (VII) and hydroxymethyltriazoloendralazine (VIII)] in human plasma was developed. After conversion of I and its internal standard to triazolopyridopyridazine derivatives the latter and metabolites were separated by high-performance liquid chromatography and detected using their fluorescence. The limits of detection of the assay were 1 nmol/l for I and VII and 0.1 nmol/l for VIII. Intra-assay coefficients of variation were 2.5-5.1% for I (range 1000-10 nmol/l), 4.2-4.5% for VII (range 100-5 nmol/l) and 3.4-5.7% for VIII (range 100-1 nmol/l). Following oral administration of 5 and 10 mg of I to two normal volunteers (slow acetylators) peak plasma levels of I occurred between 0.75 and 1 h after the dose, and declined in a biexponential fashion. The terminal half-life ranged from 2.8-3.7 h. These results contrast with those obtained for hydralazine in plasma where in vitro and in vivo half-lives were less than 30 min.