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The aim of this study was to determine the optimal biological sample and the optimal extraction technique for monitoring methadone concentrations in biological samples. We analysed methadone in serum and whole blood samples using gas chromatography/mass spectrometry (GC/MS). Before analysis, we compared five solid-phase extraction (SPE) and two liquid-liquid extraction (LLE) methods and determined that SPE with Supelco LC-18 in serum yielded the best extraction efficiency. The limit of detection was 10 ng mL-1 and the limit of quantification 25 ng mL-1. Correlation coefficient was over 0.999 for the methadone calibration curve in linear range from 50 to 2000 ng mL-1. Intra and inter-day accuracy and precision of the method was satisfactory. The method was successfully applied for determining serum methadone in patients on maintenance therapy.
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