Simple high-performance liquid chromatographic method for the determination of a new phytochemical drug, fellavine, and its metabolites in human and rat plasma and urine

Abstract The use of a small precolumn instead of an injection loop for the determination of a new phytochemical drug, fellavine, and its metabolites is described. The method combines the direct injection of plasma and urine into the reversed-phase precolumn with separation on a Spheri-5 RP-18 analytical column. Different sorbents in the precolumn were compared. A recovery of fellavine and its metabolites from biological fluids except rat plasma of almost 100% was achieved on Chrompack RP (30–40 μm) and LiChrosorb RP-18 (7 μm). For rat plasma only the last sorbent gave 80% fellavine recovery. The influence of the protein binding on the fellavine recovery was examined. The limit of detection was equal to 0.05 μg/ml fellavine for plasma and 0.02 μg/ml for urine. To enhance the limit of detection longer precolumns were preferred.