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

• Published in: Journal of pharmaceutical and biomedical analysis 1998-11, Vol.18 (3), p.403-409
• Main Authors: Lin, Ge, Nnane, Ivo P, Damani, Lyaquatali A, Tse, Kai K, Chow, Albert H.L, Kadaba, Pankaja K
• Format: Article
• Language: English
• Subjects: AAP-Cl; Aminoalkylpyridine anticonvulsant; Analysis; Aniline Compounds - analysis; Aniline Compounds - blood; Aniline Compounds - metabolism; Aniline Compounds - urine; Animals; Anticonvulsants - analysis; Anticonvulsants - blood; Anticonvulsants - metabolism; Anticonvulsants - urine; Biological and medical sciences; Calibration; Chromatography, High Pressure Liquid - methods; Drug Stability; General pharmacology; HPLC analysis; Medical sciences; Pharmacology. Drug treatments; Pyridines - analysis; Pyridines - blood; Pyridines - metabolism; Pyridines - urine; Rats; Reproducibility of Results; Sensitivity and Specificity; Ultraviolet Rays
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(98)00097-1

A simple, rapid and specific high performance liquid chromatographic (HPLC) method for the quantitation of 2-(4-chlorophenyl)amino-2-(4-pyridyl)ethane (AAP-Cl) and identification of its putative metabolite, 2-(4-chlorophenyl)amino-2-(4-pyridyl)ethanol ( β-AA) in rat blood and urine has been developed. AAP-Cl, β-AA and an appropriate internal standard were extracted from rat biofluids by a solid phase extraction technique using C18 cartridges prior to the HPLC analysis. The extractibility was 92% for AAP-Cl and 98% for β-AA. The HPLC analysis employed a symmetrical or standard reversed-phase HPLC column (Apex ODS, 5 μm, 25 cm×0.46 cm) for blood or urine analysis, a mobile phase of water–methanol–acetonitrile (40:30:30) containing 20 μl 100 ml −1 diethylamine at a flow rate of 1 ml min −1, and UV detection 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-Cl in rat biofluids were shown to be linear in both low and high concentration ranges (blood: 0–1 and 1–10 μg ml −1; urine: 0–10 and 10–100 μg ml −1) with intra- and inter-day coefficients of variation of no more than 18% for blood and 14% for urine. The method developed was successfully applied to a preliminary analysis of intact AAP-Cl in both blood and urine obtained from rats dosed with AAP-Cl.