Abstract
While the molecular mechanisms underlying postmortem change have been exhaustively investigated, the establishment of an objective and reliable means for estimating postmortem interval (PMI) remains an elusive feat. In the present study, we exploit low molecular weight metabolites to estimate postmortem interval in mice. After sacrifice, serum and muscle samples were procured from C57BL/6J mice (n = 52) at seven predetermined postmortem intervals (0, 1, 3, 6, 12, 24, and 48 h). After extraction and isolation, low molecular weight metabolites were measured via gas chromatography/mass spectrometry (GC/MS) and examined via semi-quantification studies. Then, PMI prediction models were generated for each of the 175 and 163 metabolites identified in muscle and serum, respectively, using a non-linear least squares curve fitting program. A PMI estimation panel for muscle and serum was then erected which consisted of 17 (9.7 %) and 14 (8.5 %) of the best PMI biomarkers identified in muscle and serum profiles demonstrating statistically significant correlations between metabolite quantity and PMI. Using a single-blinded assessment, we carried out validation studies on the PMI estimation panels. Mean ± standard deviation for accuracy of muscle and serum PMI prediction panels was −0.27 ± 2.88 and −0.89 ± 2.31 h, respectively. Ultimately, these studies elucidate the utility of metabolomic profiling in PMI estimation and pave the path toward biochemical profiling studies involving human samples.
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Acknowledgments
The authors wish to express their gratitude to Ms. Azumi Kuse (Department of Legal Medicine, Kobe University Graduate School of Medicine) and Dr. Koji Yamamoto for their valuable technical contributions.
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All of the animal experiments performed in this study were approved by the institutional animal care and use committee and carried out according to the Kobe University Animal Experimentation Regulations. Specimens collected for subsequent analysis were utilized in accordance with the guidelines set forth by the Kobe University Graduate School of Medicine.
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Kaszynski, R.H., Nishiumi, S., Azuma, T. et al. Postmortem interval estimation: a novel approach utilizing gas chromatography/mass spectrometry-based biochemical profiling. Anal Bioanal Chem 408, 3103–3112 (2016). https://doi.org/10.1007/s00216-016-9355-9
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DOI: https://doi.org/10.1007/s00216-016-9355-9