KU Leuven Toxicology and Pharmacology, Campus Gasthuisberg, Onderwijs en Navorsing 2, Herestraat 49, PO Box 922, 3000 Leuven, Belgium.
KU Leuven, Imaging and Pathology Department, Division Forensic Biomedical Sciences, Campus Sint-Rafaël, Kapucijnenvoer 33, 3000 Leuven, Belgium.
Forensic Sci Int. 2020 Apr;309:110225. doi: 10.1016/j.forsciint.2020.110225. Epub 2020 Feb 29.
Blood analysis is the golden standard in the field of forensic toxicology. However, when extended decomposition of the remains has occurred, alternative matrices are required. Skeletal tissue may provide an appropriate sample of choice since it is very resistant to putrefaction. However, today, the absence of reference data of drug concentrations in skeletal tissue poses a problem to meaningfully and reliably conduct toxicological testing on human skeletal material. The present study investigates the viability of skeletal tissue as an alternative matrix to evaluate xenobiotic consumption in legal cases. Blood, bone tissue and bone marrow of different forensic cases were screened for 415 compounds of forensic interest. Afterwards, methadone, clomipramine, citalopram and their respectively metabolites positive samples were quantified using fully validated methods. Sample preparation was carried out by SPE (whole blood and bone marrow), methanol extraction (bone sections) or protein precipitation (whole blood). All samples were analyzed using liquid chromatography coupled to a triple quad mass spectrometer. Multiple drugs were successfully identified in all sampled matrices. In bone (marrow) not as many substances were detected as in blood but it poses a valid alternative when blood is not available. Especially bone marrow showed big potential with a concordance of 80.5% with blood. Clomipramine, citalopram and their metabolites were proven to be detectable and quantifiable in all specimens sampled. Bone marrow showed the highest concentrations followed by blood and bone tissue. When citalopram blood concentrations were correlated with the bone concentrations, a linear trend could be detected. The same was seen between blood and bone marrow for citalopram concentrations. Methadone was also proven to be detectable in all specimens sampled. However, its metabolites EMDP and EDPP were absent or below the LOD in some samples. Overall, methadone concentrations were higher in bone marrow than in bone. With exception of one case, blood concentrations were higher than bone concentrations. For methadone, a linear trend could be found between blood and bone concentration. Comparing methadone concentrations in blood and bone marrow an exponential trend could be seen. In conclusion, these findings show the potential forensic value of bone and bone marrow as an alternative matrix. Aside to that, a standard protocol for the sample collection and processing is proposed.
血液分析是法医毒理学领域的金标准。然而，当遗体的分解程度较高时，就需要使用其他基质。骨骼组织可能是一种合适的选择，因为它非常耐腐败。然而，目前，骨骼组织中药物浓度的参考数据缺失，这给对人类骨骼材料进行有意义和可靠的毒理学测试带来了问题。本研究旨在探讨骨骼组织作为替代基质评估法医案例中外源物质消耗的可行性。对不同法医案例的血液、骨组织和骨髓进行了 415 种法医感兴趣化合物的筛查。随后，采用完全验证的方法对检出的美沙酮、氯米帕明、西酞普兰及其代谢物阳性样本进行定量分析。采用 SPE（全血和骨髓）、甲醇提取（骨切片）或蛋白沉淀（全血）进行样品制备。所有样品均采用液相色谱-三重四极杆质谱联用进行分析。在所有采样基质中均成功鉴定出多种药物。在骨骼（骨髓）中检测到的物质没有全血中那么多，但在没有全血时，它是一种有效的替代物。尤其是骨髓，其与全血的一致性高达 80.5%，具有很大的潜力。氯米帕明、西酞普兰及其代谢物在所有采集的标本中均被证明是可检测和定量的。骨髓中的浓度最高，其次是血液和骨组织。当西酞普兰的血液浓度与骨浓度相关时，可以检测到线性趋势。在血液和骨髓中也可以看到西酞普兰浓度的这种关系。美沙酮也被证明在所有采集的标本中均是可检测的。然而，在一些样本中，其代谢物 EMDP 和 EDPP 缺失或低于检测限。总体而言，骨髓中的美沙酮浓度高于骨组织。除了一个案例外，血液中的浓度都高于骨组织。对于美沙酮，在血液和骨浓度之间可以发现线性趋势。比较血液和骨髓中美沙酮浓度时，可以看到指数趋势。综上所述，这些发现表明骨骼和骨髓作为替代基质具有潜在的法医学价值。除此之外，还提出了一种用于样本采集和处理的标准方案。
