Skeletal tissue, a viable option in forensic toxicology? A view into post mortem cases

Vandenbosch, Michiel; Rooseleers, Lukas; Bogaert, Wouter Van Den; Wuestenbergs, Joke; Voorde, Wim Van de; Cuypers, Eva

doi:10.1016/j.forsciint.2020.110225

Cited by 13 publications

(11 citation statements)

References 35 publications

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“…Detection of methadone and metabolite in postmortem forensic human bone A clavicular bone of a 37-year-old male who died of an overdose of methadone was sectioned crosssectional and imaged as described. Blood concentrations of methadone and EDDP were respectively 3727.4 ng/mL and 32.9 ng/mL as described in Vandenbosch et al [25]. When looking at the distribution image of methadone, the bone marrow could be distinguished from the surrounding bone (see Fig.…”

Section: Targeted Detection Of Methadone and Eddpmentioning

confidence: 97%

Sample preparation of bone tissue for MALDI-MSI for forensic and (pre)clinical applications

et al. 2020

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In the past decades, matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) has been applied to a broad range of biological samples, e.g., forensics and preclinical samples. The use of MALDI-MSI for the analysis of bone tissue has been limited due to the insulating properties of the material but more importantly the absence of a proper sample preparation protocol for undecalcified bone tissue. Undecalcified sections are preferred to retain sample integrity as much as possible or to study the tissue-bone bio interface in particular. Here, we optimized the sample preparation protocol of undecalcified bone samples, aimed at both targeted and untargeted applications for forensic and preclinical applications, respectively. Different concentrations of gelatin and carboxymethyl cellulose (CMC) were tested as embedding materials. The composition of 20% gelatin and 7.5% CMC showed to support the tissue best while sectioning. Bone tissue has to be sectioned with a tungsten carbide knife in a longitudinal fashion, while the sections need to be supported with double-sided tapes to maintain the morphology of the tissue. The developed sectioning method was shown to be applicable on rat and mouse as well as human bone samples. Targeted (methadone and EDDP) as well as untargeted (unknown lipids) detection was demonstrated. DHB proved to be the most suitable matrix for the detection of methadone and EDDP in positive ion mode. The limit of detection (LOD) is estimated to approximately 50 pg/spot on bone tissue. The protocol was successfully applied to detect the presence of methadone and EDDP in a dosed rat femur and a dosed human clavicle. The best matrices for the untargeted detection of unknown lipids in mouse hind legs in positive ion mode were CHCA and DHB based on the number of tissue-specific peaks and signal-to-noise ratios. The developed and optimized sample preparation method, applicable on animal and human bones, opens the door for future forensic and (pre)clinical investigations.

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Section: Targeted Detection Of Methadone and Eddpmentioning

confidence: 97%

Sample preparation of bone tissue for MALDI-MSI for forensic and (pre)clinical applications

et al. 2020

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“…One of the possible reasons for the lack of research on bone (marrow) is the difficulty in obtaining bones from cases. In a previous project, a linear trend could be found in human autopsy samples between blood and bone concentration for methadone ( 21 ). When methadone concentrations in blood and BM were compared, an exponential trend could be seen.…”

Section: Introductionmentioning

confidence: 90%

“…Cases were selected after a positive screening result for tramadol, morphine, fentanyl, codeine, cocaine or any of their metabolites in blood using the method as described by Vandenbosch et al. ( 21 ). Twenty-two cases were selected.…”

Section: Methodsmentioning

confidence: 99%

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Postmortem Analysis of Opioids and Metabolites in Skeletal Tissue

Vandenbosch

Pajk

Bogaert

et al. 2021

Journal of Analytical Toxicology

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Every year, thousands of suspicious deaths are accounted for by an overdose of opioids. Occasionally all traditional matrices are unavailable due to decomposition. Skeletal tissue may pose a valid alternative. However, reference data on post-mortem concentrations in bone tissue and bone marrow is sparse. Therefore, a liquid chromatography tandem mass spectrometry method was developed and fully validated for the analysis of 4 opioids and 2 metabolites (tramadol, O-desmethyltramadol, morphine, fentanyl, norfentanyl, codeine) in bone tissue and bone marrow. Sample preparation was performed using solid phase extraction (bone marrow), methanolic extraction (bone) and a protein precipitation (whole blood). All validation parameters were successfully fulfilled. This method was applied to analyze 22 forensic cases involving opioids. All 6 opioids were proven to be detectable and quantifiable in all specimens sampled. When tramadol blood concentrations were correlated with bone concentrations, a linear trend could be detected. The same was seen between tramadol blood and bone marrow concentration. A similar linear trend was seen when correlating codeine blood concentration with bone and bone marrow concentration. Although some variability was detected, the same linear trend was seen for morphine. For fentanyl and norfentanyl, the sample size was too small to draw conclusions, regarding correlation. As far as the authors know this is the first-time fentanyl and norfentanyl are quantified in skeletal tissue. In conclusion, due to the absence of reference data for drugs in skeletal tissue, these findings are a step forward towards a more thorough understanding of drug concentration found in post-mortem skeletal tissue.

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“…Its placement protects it from contamination as the bone remains intact. Furthermore, in the case of corpses in an advanced stage of decomposition or skeletonization, it is often one of the few materials available, due to its protection against the influence of fungi and bacteria, which delays the decay process [18][19][20].…”

Section: Introductionmentioning

confidence: 99%

The DI-SPME Method for Determination of Selected Narcotics and Their Metabolites, and Application to Bone Marrow and Whole Blood Analysis

et al. 2022

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The present investigation utilised the quick and easy SPME/LC-MS method to determine selected narcotic substances and their metabolites in whole blood. The study included qualitative analysis and validation of the method. Analytes were determined in the linearity range of 25–300 ng/mL. The precision during and between days (in general CV < 13.41%), and the LOD which results in between 0.36 and 11.08 ng/mL, and the LOQ between 1.20 and 36.90 ng/mL were investigated. The validation results obtained, as well as the results of subsequent in-laboratory tests, confirmed the applicability of the method in the analysis of blood samples. An attempt to apply the method to the analysis of bone marrow samples has yielded promising results; however, more detailed studies are needed in this area.

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Skeletal tissue, a viable option in forensic toxicology? A view into post mortem cases

Cited by 13 publications

References 35 publications

Sample preparation of bone tissue for MALDI-MSI for forensic and (pre)clinical applications

Sample preparation of bone tissue for MALDI-MSI for forensic and (pre)clinical applications

Postmortem Analysis of Opioids and Metabolites in Skeletal Tissue

The DI-SPME Method for Determination of Selected Narcotics and Their Metabolites, and Application to Bone Marrow and Whole Blood Analysis

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