This study analyzed whole blood from five case samples of confirmed multiple-drug overdose previously screened and quantified by the West Virginia Office of the Chief Medical Examiner, with the current study using a direct injection multi-stage mass spectrometric (MS) method to confirm the identity of 26 analytes at or above 10 ng/ml, using 16 deuterium-labeled internal standards.
Samples were spiked with internal standards, precipitated with acetonitrile, and centrifuged. Samples were further diluted with either 0.1 percent formic acid or 0.1 percent ammonium hydroxide in methanol prior to injection into an electrospray ionization ion trap mass spectrometer (MS). Ions were monitored as MS-MS or MS3 product ions. In all cases, analysis by MS-MS confirmed the presence of the drugs and metabolites when the internal standards were detected. Detection of characteristic MS3 ions was used for further confirmation of the presence of parent drugs in all but three instances. Total analysis time was less than 1 h. Although only useful for qualitative or confirmatory purposes, this direct injection MSn method provides a simple and rapid confirmation of multiple drugs that have been previously identified and quantified by gas chromatographic-MS or liquid chromatographic-MS analytical methods. (publisher abstract modified)