, Volume 48, Issue 7–8, pp 555–560 | Cite as

High-performance liquid chromatographic assay for simultaneous determination of tramadol and its active metabolite in human plasma. Application to pharmacokinetic studies

  • M. A. Campanero
  • B. Calahorra
  • E. García-Quetglás
  • M. Escolar
  • J. Honorato


A sensitive liquid chromatographic assay for the quantitative determination of the opioid analgesic tramadol and its active metabolite is described. Fluconazole was used as internal standard. The assay involved a singletert-butyl methyl ether extraction and LC analysis with fluorescence detection. Chromatography was at 30°C pumping an isocratic mobile phase of acetonitrile-water (19∶81, v/v) containing 0.06M NaH2PO4 and 0.05M triethylamine, adjusted to pH 7.90, at 1 mL min−1 through a reversed-phase, 250×4 mm base-stable column. The limit of quantitation of tramadol and its active metabolite was 1 ng mL−1, only 0.5 mL plasma sample was required for the determination. The calibration curve was linear from 1–1000 ng mL−1. Intra and inter-day precision (C.V.) did not exceed 10%. Mean recoveries of 96.38% for tramadol and 96.62% forO-demethyltramadol with CVs of 0.43% and 1.46% were obtained. Applicability of the method was demonstrated by a pharmacokinetic study on normal volunteers who received 100 mg tramadol intravenously.

Key Words

Column liquid chromatography Tramadol andO-demethyl tramadol in plasma Pharmacokinetics 


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Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1998

Authors and Affiliations

  • M. A. Campanero
    • 1
  • B. Calahorra
    • 1
  • E. García-Quetglás
    • 1
  • M. Escolar
    • 1
  • J. Honorato
    • 1
  1. 1.Servicio de Farmacología Clínica, Clínica Universitiria de navarraUniversidad de NavarraPamplonaSpain

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