Sensitive microanalysis of gabapentin by high-performance liquid chromatography in human serum using pre-column derivatization with 4-chloro-7-nitrobenzofurazan: Application to a bioequivalence study

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Gholamreza Bahrami, Bahareh Mohammadi

Abstract

Most of the published methods for analysis of gabapentin, an antiepileptic agent, in human serum require automated o-phthalaldehyde derivatization of the drug and immediate injection of the unstable derivatives formed. A new, very sensitive and simple high-performance liquid chromatographic method for quantitation of the drug in human serum using 4-chloro-7-nitrobenzofurazan (NBD-Cl) as a fluorescent labeling agent is presented. In this method the sensitivity was significantly improved and the limit of quantification of 0.002 microg/ml was obtained using 100 microl serum sample and 10 microl injection. However, the LOQ can be improved by increasing the sampling volume. The procedure involved protein precipitation of serum by acetonitrile followed by derivatization with NBD-Cl. Amlodipine was used as internal standard and chromatographic separation was performed on a Shimpack CLC-C18 (150 mm x 4.6 mm) column. The fluorescence derivative of the drug was monitored at excitation and emission wavelengths of 470 and 537 nm, respectively. A mobile phase consisting of methanol and sodium phosphate buffer (0.05 M; pH 2.5) containing 1 ml/l triethylamine (65:35, v/v) was used. The calibration curve was linear over the concentration range...Continue Reading

References

Nov 15, 2000·Journal of Chromatography. B, Biomedical Sciences and Applications·D F CholletG Anderegg
Sep 24, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·T A C Vermeij, P M Edelbroek
Mar 8, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Daniëlle C R BorreyMichel R Langlois

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Citations

Apr 7, 2010·Journal of Pharmaceutical and Biomedical Analysis·Laura MercoliniMaria Augusta Raggi
May 23, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Hassan JalalizadehAlireza Jahangiri
Jan 2, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Gholamreza Bahrami, Bahareh Mohammadi
Jul 11, 2008·Journal of Cellular Biochemistry·Seyeon ParkYong-Woo Hong
Mar 1, 2008·Journal of Veterinary Pharmacology and Therapeutics·L DirikoluT Tobin

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