Synthesis of d1-N-ethyltramadol as an internal standard for the quantitative determination of tramadol in human plasma by gas chromatography-mass spectrometry

A gas chromatography-mass spectrometry (GC-MS) assay for the determination of tramadol in human plasma is presented. The synthesis of an N-ethyl analogue of the drug is described and its use as an internal standard for the quantitative measurement of tramadol in human plasma is described. The method involves extraction at plasma pH and analysis of the underivatized drug by gas chromatography-electron ionization mass spectrometry using m/z 58 and 73 for detection of tramadol and internal standard, respectively. The calibration curve was linear in the range of 5-640 ng/ml plasma (r = 0.9999). The method was validated in the abovementioned calibration range. Data on solution stability, long- and short-term stability of tramadol in plasma samples, freeze-thaw-stability, as well as inter- and intra-day precision and accuracy have been evaluated and are presented. The application of the method to the pharmacokinetic profiling of the drug is demonstrated. (C) 2004 Elsevier B.V. All rights reserved.