In order to characterize the protein binding of a drug, it is necessary to have a method which is close to in vivo conditions and fast in the course of measurement. The continuous ultrafiltration fulfils both requirements for substances with a high extent of protein binding. In this study, 18 gyrase inhibitors in clinical practice, characterized by a lower extent of protein binding, were subjected to the titration procedure of the continuous ultrafiltration using bovine and human serum albumin (BSA, HSA), and human plasma. The results of the continuous ultrafiltration were found to be similar to those obtained by means of the 'classical' discontinuous ultrafiltration using plasma (correlation between continuous and discontinuous ultrafiltration r2 = 0.87). In the cases of pipemidic acid, enoxacin and rufloxacin, the continuous method gave approximately 20% lower degrees of protein binding than the discontinuous procedure, which utilizes plasma having the full range of proteins. It is likely that these drugs bind mainly to other proteins in plasma than HSA. This finding proves that this fast method is worthwhile in the whole range of protein binding.