High performance liquid chromatographic analysis of a novel aminoalkylpyridine anticonvulsant 2-(4-chlorophenyl)amino-2-(4-pyridyl)ethane

A simple, rapid and specific high performance liquid chromatographic (HPLC) method for the quantitation of 2-(4-chlorophenyl)amino-2-(4-pyridyl)ethane (AAP-CI) and identification of its putative metabolite, 2-(4-chlorophenyl) amino-2-(4-pyridyl)ethanol (beta-AA) in rat blood and urine has been develo ped. AAP-CI, beta-AA and an appropriate internal standard were extracted fr om rat biofluids by a solid phase extraction technique using Cls cartridges prior to the HPLC analysis. The extractibility was 92% for AAP-Cl and 98% for beta-AA. The HPLC analysis employed a symmetrical or standard reversed- phase HPLC column (Apex ODS, 5 mu m, 25 cm x 0.46 cm) for blood or urine an alysis, a mobile phase of water-methanol-acetonitrile (40:30:30) containing 20 mu l 100 ml(-1) diethylamine at a Bow rate of 1 mi min(-1), and UV dete ction at 254 nm. The limit of detection was 100 ng ml(-1) for both analytes in both blood and urine. The calibration curves for AAP-CI in rat biofluid s were shown to be linear in both low and high concentration ranges (brood: 0-1 and 1-10 mu g ml(-1); urine: 0-10 and 10-100 mu g ml(-1)) with intra- and inter-day coefficients of variation of no more than 18% for blood and 1 4% for urine. The method developed was successfully applied to a preliminar y analysis of intact AAP-CI in both blood and urine obtained from rats dose d with AAP-CI. (C) 1998 Elsevier Science B.V. All rights reserved.