Simultaneous Determination of Protein-Free and Total Positron Emission Tomography Radioligand Concentrations in Plasma Using High-Performance Frontal Analysis Followed by Mixed Micellar Liquid Chromatography: Application to [11C]PBR28 in Human Plasma

A two-dimensional liquid chromatographic (LC) system was developed for the determination of protein-free and total (free + bound forms) positron emission tomography (PET) radioligand concentrations in plasma by direct plasma injection. The unbound PET radioligand was first analyzed by high-performance frontal analysis using a short gel-filtration column and phosphate buffered saline solution as the mobile phase. All the collected effluent from the gel-filtration column was then transferred to the second dimension consisting of a monolithic C-18 column and mixed (anionic and nonionic surfactants) micellar eluent for determination of the total PET radioligand concentration. The simultaneous analysis of protein binding and radiometabolism of [11C]PBR28 was completed within 11 min without any pretreatment of plasma and employing a single analytical system. This system allowed highly sensitive analysis of total [11C]PBR28 with a limit of detection (LOD) of 1.6 becquerel (Bq). The LOD for the determination of u...