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-Cl) and identification of its putative metabolite, 2-(4-chlorophenyl)amino-2-(4-pyridyl)ethanol (beta-AA) in rat blood and urine has been developed. AAP-Cl, beta-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 beta-AA. The HPLC analysis employed a symmetrical or standard reversed-phase HPLC column (Apex ODS, 5 microm, 25 cm x 0.46 cm) for blood or urine analysis, a mobile phase of water methanol acetonitrile (40:30:30) containing 20 microl 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 microg ml(-1); urine: 0-10 and 10-100 microg 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.
Lin
G
G
Department of Pharmacology, The Chinese University of Hong Kong, Shatin, NT. linge@cuhk.edu.hk
Nnane
I P
IP
Damani
L A
LA
Tse
K K
KK
Chow
A H
AH
Kadaba
P K
PK
D000814:Aniline Compounds / Q000032:analysis* / Q000097:blood / Q000378:metabolism / Q000652:urine; D000818:Animals; D000927:Anticonvulsants / Q000032:analysis* / Q000097:blood / Q000378:metabolism / Q000652:urine; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D011725:Pyridines / Q000032:analysis* / Q000097:blood / Q000378:metabolism / Q000652:urine; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014466:Ultraviolet Rays
C119418:2-(4-chlorophenyl)amino-2-(4-pyridyl)ethane; D000814:Aniline Compounds; D000927:Anticonvulsants; D011725:Pyridines