Using porcine bone marrow to analyze fenproporex and its metabolite amphetamine for forensic toxicological purposes: method development and validation

The possibility of detecting drugs of abuse in bone marrow represents an important issue in forensic toxicology. Bone marrow is an alternative matrix for cases in which the matrices commonly used in forensic practice are unavailable or unfeasible for analysis. The aim of this study was to develop a method for bone cleaning and bone marrow pretreatment and subsequently validate a quantitative method for the simultaneous analysis of fenproporex and amphetamine in porcine bone marrow. Bones were cleaned with proteolytic enzymes and then were cut into small fragments in order to remove the bone marrow. A homogenate was obtained after adding an acidic solution to the bone marrow. Then, the bone marrow homogenate was cleaned with ethyl ether and, after pH adjustment, extracted by liquid-liquid extraction (LLE) with ethyl ether. The dry residue obtained after solvent evaporation was derivatized with N -methyl-bis-trifluoroacetamide (MBTFA)/ N -methyl- N -(trimethylsilyl)trifluoroacetamide (MSTFA) and then analyzed by gas chromatography-mass spectrometry (GC-MS). The developed method allowed the identification and quantification of target analytes in the studied matrix. The validation procedure showed good linearity ( r 2 > 0.99) and recovery (>80%) for fenproporex and amphetamine. The limits of detection and quantitation were 1 ng mg −1 and 5 ng mg −1 , respectively, for both analytes. The obtained results are of great importance and may be suitable for potential use in forensic practice. A new method for bone cleaning and bone marrow pretreatment was developed for application in forensic toxicological routine.