Determination of drugs of abuse and their metabolites in human plasma by liquid chromatography-mass spectrometry. An application to 156 road fatalities

A method, using 0.2 ml of plasma, was designed for the simultaneous determination of morphine, 6-monoacetylmorphine, amphetamine, methamphetamine, MDA, MDMA, MDEA, MBDB, benzoylecgonine and cocaine. The drugs were analysed by LC-MS, after solid phase extraction in the presence of the deuterated analogues. Reversed phase separation on an Atlantis dC18 column was achieved in 10 min, under gradient conditions. The method was full validated, including linearity (2-250 ng/ml, r2>0.99), recovery (>50%), within-day and between-day precision and accuracy (CV and bias <15%), limit of detection (0.5 and 1 ng/ml) and quantitation (2 ng/ml), relative ion intensities and no matrix effect was observed. The procedure showed to be sensitive and specific, and was applied to 156 real cases from road fatalities (7.1% cases positive to cocaine and 0.6% to designer drugs).