Human bone marrow as a tissue in post-mortem identification and determination of psychoactive Substances-Screening methodology

•A MAE/UHPLC-MS method for identification of psychoactive drugs in human bone marrow was developed.•Lipids removal was incorporated as effective means of sample preparation for complex matrices.•Optimum extraction conditions were found using modified simplex method.•The developed method was applied to the analysis of samples from three real forensic cases.

The aim of this study was to apply a new instrumental approach to the analysis of human bone marrow for forensic purposes. A new screening method for the detection of more than 30 psychoactive drugs in bone marrow was developed and applied to case samples. The drugs used in this study belonged to the following groups: benzodiazepines (BZDs, n = 16), tricyclic antidepressants (TCAs, n = 5), selective serotonin reuptake inhibitors (SSRI, n = 4), serotonin-norepinephrine reuptake inhibitors (n = 1), anticonvulsants (n = 1), imidazopyridines (n = 1) and piperazines (n = 3). The sample preparation procedure involved microwave-assisted extraction (MAE) and the experimental settings were optimized using the simplex method. Separation and detection was carried out using a UHPLC-TOF system. The method were validated using marrow samples, and further applied in the analysis of three case samples, in which diazepam, nordiazepam, citalopram, doxepin and paroxetine were successfully detected. Finally the presented method is a good example of an assay that could potentially find an application in forensic analysis.