Comparative use of aqueous humour 1 H NMR metabolomics and potassium concentration for PMI estimation in an animal model

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ORIGINAL ARTICLE

Comparative use of aqueous humour 1H NMR metabolomics and potassium concentration for PMI estimation in an animal model Emanuela Locci 1 & Matteo Stocchero 2 & Rossella Gottardo 3 & Fabio De-Giorgio 4,5 Matteo Nioi 1 & Alberto Chighine 1 & Franco Tagliaro 3,6 & Ernesto d’Aloja 1

&

Roberto Demontis 1

&

Received: 15 September 2020 / Accepted: 12 November 2020 # The Author(s) 2020

Abstract Estimation of the post-mortem interval (PMI) remains a matter of concern in the forensic scenario. Traditional and novel approaches are not yet able to fully address this issue, which relies on complex biological phenomena triggered by death. For this purpose, eye compartments may be chosen for experimental studies because they are more resistant to post-mortem modifications. Vitreous humour, in particular, has been extensively investigated, with potassium concentration ([K+]) being the marker that is better correlated with PMI estimation. Recently, a 1H nuclear magnetic resonance (NMR) metabolomic approach based on aqueous humour (AH) from an animal model was proposed for PMI estimation, resulting in a robust and validated regression model. Here we studied the variation in [K+] in the same experimental setup. [K+] was determined through capillary ion analysis (CIA) and a regression analysis was performed. Moreover, it was investigated whether the PMI information related to potassium could improve the metabolome predictive power in estimating the PMI. Interestingly, we found that a part of the metabolomic profile is able to explain most of the information carried by potassium, suggesting that the rise in both potassium and metabolite concentrations relies on a similar biological mechanism. In the first 24-h PMI window, the AH metabolomic profile shows greater predictive power than [K+] behaviour, suggesting its potential use as an additional tool for estimating the time since death. Keywords Aqueous humour . Animal model . PMI . Potassium concentration . CIA . 1H NMR metabolomics

Introduction Accurate post-mortem interval (PMI) estimation is a challenging issue in forensic pathology. Traditional biochemical * Emanuela Locci [email protected] 1

Department of Medical Sciences and Public Health, Section of Legal Medicine, University of Cagliari, Cittadella Universitaria di Monserrato, 09042 Monserrato, CA, Italy

2

Department of Women’s and Children’s Health, University of Padova, Padova, Italy

3

Department of Diagnostics and Public Health, Unit of Forensic Medicine, University of Verona, Verona, Italy

4

Department of Health Surveillance and Bioethics, Section of Legal Medicine, Catholic University of Rome, Rome, Italy

5

Fondazione Policlinico Universitario A. Gemelli IRCCS , Rome, Italy

6

Institute of Pharmacy and Translational Medicine, Sechenov First Medical University, Moscow, Russia

methods have been applied to identify metabolite changes after death in several biofluids [1–3]. Vitreous humour (VH) is the most studied fluid for post-mortem analysis given its correlation with ante-mortem serum compo