Furosemide binding by human albumin: comparison of two methods of fluorescence quenching analysis

Disagreement exists over the primary-site binding constant for the interaction of furosemide with human albumin. Disagreement also exists over which experimental methods are accurate in this particular interaction. Therefore, furosemide binding by human albumin was examined using albumin fluorescence quenching by both the method of Levine and the method of Steiner et al. The binding constants obtained by each method differed greatly, with the results of the latter method being similar to those of other experimental methods. It was concluded that the method of Levine overestimates the binding constant for this drug-protein interaction.