DIACETYLMORPHINE AND ITS METABOLITES IN PLASMA BY HPLC WITH DIODE-ARRAY AND ATMOSPHERIC PRESSURE IONIZATION MASS SPECTROMETRIC DETECTION

Abstract

Based on reversed phase high performance liquid chromatography with gradient elution and diode-array detection (HPLC-DAD) a specific and sensitive assay for the quantitation of diacetylmorphine (heroin, DAM) and its primary and secondary metabolites 6-acetylmorphine, morphine, morphine-3-β-D-glucuronide, morphine-6-β-D-glucuronide and nor-morphine in plasma was developed. Confirmation of the peak assignment was performed by atmospheric pressure ionization-mass spectrometry (API-MS). Plasma samples were extracted on C18 ODS-2 columns using an automated solid-phase extraction (SPE) unit. The recovery was between 45 and 68%. The quantitation limit of DAM and its metabolites was 50 and 10 ng/mL, respectively. The inter-day precision was in the range of 4.0 to 17.6%. The described method was applied for pharmacokinetic studies which were part of the evaluation and validation of DAM preparations used within a heroin-assisted treatment program for dependent drug users.