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A sensitive and efficient method was developed for determination of tramadol and its metabolite ( O -desmethyltramadol) in human urine by gas chromatography–mass spectrometry. Tramadol,  O -desmethyltramadol and medazepam (internal standard) were extracted from human urine with a mixture of ethylacetate and diethylether mixture (1 : 1, v/v) at basic pH with liquid–liquid extraction. The calibration curves were linear ( r  = 0.99) over tramadol and  O -desmethyltramadol concentrations ranging from 10 to 200 ng/mL and 7.5 to 300 ng/mL, respectively. The method had an accuracy of &gt;95% and intra- and interday precision (relative standard deviation %) of ≤4.93 and ≤4.62% for tramadol and  O -desmethyltramadol, respectively. The extraction recoveries were found to be 94.1 ± 2.91 and 96.3 ± 3.46% for tramadol and  O -desmethyltramadol, respectively. The limit of quantification using 0.5 mL human urine was 10 ng/mL for tramadol and 7.5 ng/mL for  O -desmethyltramadol. After oral administration of 100 mg of tramadol hydrochloride to a patient, the urinary excretion was monitored during 24 h. About 15% of the dose was excreted as unchanged tramadol.

Simultaneous Determination of Tramadol and Its Metabolite in Human Urine by the Gas Chromatography–Mass Spectrometry Method