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Enantioselective separation and determination of adrafinil and modafinil on Chiralcel OJ‐H column in rat serum and urine using solid‐phase extraction followed by HPLC
Author(s) -
Rao R. Nageswara,
Shinde Dhananjay D.
Publication year - 2009
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1190
Subject(s) - chromatography , chemistry , metabolite , elution , solid phase extraction , urine , enantiomer , detection limit , extraction (chemistry) , high performance liquid chromatography , pharmacokinetics , pharmacology , biochemistry , medicine , organic chemistry
A simple and rapid normal‐phase HPLC method for enantiospecific separation of a psychostimulant, adrafinil (ADL), and its metabolite modafinil (MDL) in rat serum and urine was developed. The separation was accomplished on a normal‐phase polysaccharide stationary phase Chiralcel OJ‐H using n ‐hexane–ethanol (62:38 v/v) as a mobile phase at a flow rate of 1.0 mL/min. Detection was carried out at 225 nm using a photo diode array (PDA) detector. The elution order of the enantiomers was determined by a polarimeter connected in series with the PDA. ADL and its metabolite were recovered from rat serum and urine by solid phase extraction using Oasis HLB cartridges and the mean recoveries were ≥80%. The enantiomers were eluted within 15 min without any interference from endogenous substances. The calibration curves were linear ( r 2 > 0.998) in the concentration range of 1.20–500 µg/mL for ADL and MDL. The assay was specific, accurate, precise and reproducible (intra‐ and inter‐day precisions RSDs <7.2%). ADL in rat serum was stable over three freeze–thaw cycles at ambient temperature for 4 h. The method was successfully applied to pharmacokinetic studies of adrafinil after an oral administration to rats. Copyright © 2009 John Wiley & Sons, Ltd.