Development and validation of bone extraction procedure in forensic toxicology: A comparison of different extraction techniques

Purpose: Forensic toxicology plays a crucial role in forensic sciences, focusing on the detection and interpretation of xenobiotics in various biological samples, including bone tissue. This study aimed to compare different extraction techniques for bone tissue and subsequently analyze and validate the best analytical method via liquid chromatography–tandem mass spectrometry. Methods: Samples were collected from autopsied cadavers, prepared, and subjected to four different extraction methods (EM-1 to EM-4). Results: Absolute recovery tests indicated EM-1 as the better extraction method, successfully detecting all the target analytes. Method validation for EM-1 demonstrated acceptable bias, precision, interference studies, and stability in various analytes. The validated method was then applied to ten real cases, confirming its efficacy in forensic toxicology for detecting opioids, dissociative anesthetics, stimulants, benzodiazepines, anticonvulsants, antidepressants, antipsychotic medications, and sedatives in bone tissue. Conclusions: This study validated a new extraction method for bone tissue. This technique was applied to forensic real cases.