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Monitoring of post-mortem changes of saliva N-glycosylation by nano LC/MS

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Abstract

The estimation of post-mortem interval (PMI) is a crucial part for investigations of crime and untimely deaths in forensic science. However, standard methods of PMI estimation are easily confounded by extenuating circumstances and/or environmental factors. Therefore, a panel of PMI markers obtained from a more acceptable and accurate method is necessary to definitely determine time of death. Saliva, one of the vital fluids encountered at crime scenes, contains various glycoproteins that are highly affected by biochemical environment. Here, we investigated saliva N-glycans between live and dead rats to determine the alteration of N-glycans using an animal model system because of the limitation of saliva collection from recently deceased humans. Rat saliva samples were collected both before and after death. N-Glycans were enzymatically released by PNGase F without any glycoprotein extraction. Released native glycans were purified and enriched by PGC-SPE. About 100 N-glycans were identified, profiled, and structurally elucidated by nano LC/MS and tandem MS. Sialylated N-glycans were exclusively present in abundance in live rat saliva whereas non-sialylated N-glycans including LacdiNAc disaccharides were detected in high level following death. Through in-depth investigations using quantitative comparison and statistical analysis, 14 N-glycans that significantly changed after death were identified as the potential marker candidates for PMI estimation. To the best of our knowledge, this is the first study to monitor the post-mortem changes of saliva glycosylation, with obvious forensic applications.

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Acknowledgements

The authors are grateful for the support provided by a Korea Basic Science Institute grant (C37703 for J.S.C.) and a grant from the Ministry of Science, ICT and Future Planning (NRF-2016M3A9E1918324 for H.J.A.).

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Authors and Affiliations

  1. Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Korea

    Bum Jin Kim, Chanyoung Han, Hantae Moon, Jong-Soon Choi & Hyun Joo An

  2. Asia-Pacific Glycomics Reference Site, Chungnam National University, Daejeon, Korea

    Bum Jin Kim, Chanyoung Han, Hantae Moon & Hyun Joo An

  3. Biological Disaster Analysis Team, Korea Basic Science Institute, Daejeon, Korea

    Joseph Kwon, Ik-Soon Jang & Jong-Soon Choi

  4. Forensic DNA Division, National Forensic Service, Wonju, Korea

    Si-Keun Lim & Ki-Won Park

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The authors declare that the experiments have been conducted in accordance with the protocol of the Korean Council on Animal Care and approved by the Animal Care Committee of Korea Basic Science Institute (KBSI-AEC 1715). All efforts were made to minimize animal suffering and to reduce the number of rats.

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Kim, B.J., Han, C., Moon, H. et al. Monitoring of post-mortem changes of saliva N-glycosylation by nano LC/MS. Anal Bioanal Chem 410, 45–56 (2018). https://doi.org/10.1007/s00216-017-0702-2

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