Sensitive high-performance liquid chromatographic quantitation of gabapentin in human serum using liquid-liquid extraction and pre-column derivatization with 9-fluorenylmethyl chloroformate.
Most of the published methods for analysis of gabapentin, an antiepileptic agent, in human serum are based on the same approach, involving o-phthaldialdehyde derivatization of deproteinized serum samples. The present paper however, describes a new, simple and sensitive high-performance liquid chromatographic method for determination of gabapentin in human serum using liquid-liquid extraction and 9-fluorenylmethyl chloroformate (FMOC-Cl) as pre-column labeling agent. The drug and an internal standard (azithromycin) were extracted from serum by salting-out approach using a mixture of dichloromethane-2 propanol (1:1, v/v) as the extracting solvent. The extracted analytes were subjected to derivatization with FMOC-Cl in the presence of phosphate buffer (pH 7). A mobile phase consisting of methanol-0.05 M sodium phosphate buffer (73/27, v/v; pH of 3.9) containing 1 ml/l triethylamine was eluted and chromatographic separation was performed on a Shimpack CLC-C18 (150 mm x 4.6 mm) column. The standard curve was linear over the range of 0.03-20 microg/ml and limit of quantification was 0.03 microg/ml. The performance of analysis was studied and the validated method showed excellent performance in terms of selectivity, specificity, sensitivity, precision and accuracy. No interferences were found from commonly co-administered antiepileptic agents.
Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran. gbahrami@kums.ac.ir
Gholamreza
G
Bahrami
Amir
A
Kiani
D000588:Amines / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003509:Cyclohexanecarboxylic Acids / Q000097:blood*; D004355:Drug Stability; D005449:Fluorenes / Q000737:chemistry; D000077206:Gabapentin; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D005680:gamma-Aminobutyric Acid / Q000097:blood*
D000588:Amines; D003509:Cyclohexanecarboxylic Acids; D005449:Fluorenes; D005680:gamma-Aminobutyric Acid; D000077206:Gabapentin; C054007:1-(9-fluorenyl)methyl chloroformate