The aim of the present study was to assess the suitability of different Amicon Ultra and Centrifree ultrafiltration devices for the study of the plasma protein binding process in the case of carvedilol, a highly protein-bound and lipophilic beta-blocking agent. Samples at different levels of concentration were prepared in both proteic and non-proteic matrices (human plasma, 5% human serum albumin solution and saline solution) and subjected to the classical ultrafiltration method using the different devices considered. Furthermore, an attempt to apply a previously described modified ultrafiltration method was also made. The analysis and quantification was achieved using a validated LC-MS/MS method. For the Centrifree devices, the determined unbound fractions of carvedilol and the corresponding binding degree were in accordance to literature data, while for the Amicon Ultra devices a great degree of carvedilol adsorbtion to the sample reservoir was observed, the analyte not being detected in the ultrafiltrate samples. Thus, it was further demonstrated that the type of ultrafiltration device used has a significant influence on the outcome of a plasma protein binding study. In the case of carvedilol, the evaluation of the protein binding interaction could be achieved using the Centrifree ultrafiltration devices, but not the Amicon Ultra devices.
1 Doctoral School of Medicine and Pharmacy, I.O.S.U.D., George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
2 Department of Analytical Chemistry and Drug Analysis, Faculty of Pharmacy, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania
3 Center of Advanced Medical and Pharmaceutical Research, George Emil Palade University of Medicine, Pharmacy, Science, and Technology of Targu Mures, Romania