Solid phase extraction of morphine and its metabolites from postmortem blood

A simple and rapid high-performance liquid chromatographic method is described for the determination of morphine-3-glucuronide, morphine-6-glucuronide, normorphine and morphine in postmortem blood. A solid phase extraction technique employing C18 Sep-Pak cartridges was used to recover morphine and its metabolites from 0.5 ml of blood. Reverse phase ion-pair chromatography was used to achieve separation with a C18 bonded column. The mobile phase consisted of acetonitrile, lauryl sulphate and sodium dihydrogen orthophosphate buffer at low pH. Electrochemical detection (ECD) in series with ultraviolet (UV) spectrophotometric detection (210 nm) was used for quantitation. The lower limit of detection using ECD was 10 ng/ml for all analytes and a linear response was observed to 5000 ng/ml. Coefficients of variation for all analytes ranged between 3-13% for both intra- and inter-assay. This method is reproducible, quick and easy to perform and allows morphine conjugates and morphine to be measured simultaneously in postmortem blood.