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Discovery of donor age markers from bloodstain by LC-MS/MS using a metabolic approach

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Abstract

Bloodstains are frequently encountered at crime scenes and they provide important evidence about the incident, such as information about the victim or suspect and the time of death or other events. Efforts have been made to identify the age of the bloodstain’s donor through genomic approaches, but there are some limitations, such as the availability of databases and the quality dependence of DNA. There is a need for the development of a tool that can obtain information at once from a small blood sample. The aim of this study is to identify bloodstain metabolite candidates that can be used to determine donor age. We prepared bloodstain samples and analyzed metabolites using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Eighteen molecular features (MFs) were selected as candidates using volcano plots and multivariate analysis. Based on the MS/MS spectrum of the MFs, the following nine metabolites were identified from the METaboliteLINk database: Δ2-cis eicosenoic acid, ergothioneine, adenosine 5′-monophosphate, benzaldehyde, phenacylamine, myristic acid ethyl ester, p-coumaric acid, niacinamide, and N-arachidonoyl-l-alanine. These nine age markers at high or low abundances could be used to estimate the age of a bloodstain’s donor. This study was the first to develop metabolite age markers that can be used to analyze crime scene bloodstains.

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Acknowledgements

The authors would like to thank Agilent Technologies for the technical support with constructive comments.

Funding

This research was supported and funded by the Korean National Police Agency (Project Name: Development of blood stain analysis system for scene reconstruction/Project Number: PA-I000001).

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Authors and Affiliations

  1. Department of Senior Healthcare, Graduate School, Eulji University, 712, Dongil-ro, Uijeongbu-si, Gyeonggi-do, 11759, Republic of Korea

    Hyo-Jin Kim, You-Rim Lee, Seungyeon Lee, Sohyen Kwon, Sung Hee Hyun, Ho Joong Sung & Hee-Gyoo Kang

  2. Department of Biomedical Laboratory Science, College of Health Sciences, Eulji University, Seongnam, 13135, Republic of Korea

    Yeon Tae Chun, Ho Joong Sung & Hee-Gyoo Kang

  3. Integrative Research Support Center, The Catholic University of Korea, School of Medicine, 222 Banpo-daero, Seocho-gu, Seoul, 06501, Republic of Korea

    Yeon Tae Chun

  4. Department of Biomedical Laboratory Science, College of Health Science, Eulji University, Uijeongbu, 11759, Republic of Korea

    Sung Hee Hyun & Jiyeong Lee

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  1. Hyo-Jin Kim
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  9. Hee-Gyoo Kang
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Contributions

Hyo-Jin Kim: conceptualization, methodology, formal analysis, investigation, visualization, writing—original draft preparation, writing—reviewing and editing. You-Rim Lee: methodology, investigation, visualization, writing—reviewing and editing. Seungyeon Lee: investigation. Sohyen Kwon: investigation. Yeon Tae Chun: investigation. Sung Hee Hyun: formal analysis. Ho Joong Sung: formal analysis. Jiyeong Lee: visualization, writing—reviewing and editing, supervision, project administration. Hee-Gyoo Kang: conceptualization, resources, writing—reviewing and editing, supervision, project administration, funding acquisition

Corresponding authors

Correspondence to Jiyeong Lee or Hee-Gyoo Kang.

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All participants gave written informed consent before participation in the study. This study was approved by the Institutional Review Board of Eulji Hospital (EMC 2017-03-003).

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The authors declare no competing interests.

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Supplementary Figure 1.

Experimental workflow to discover the donor age markers. (TIF 3627 kb)

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Kim, HJ., Lee, YR., Lee, S. et al. Discovery of donor age markers from bloodstain by LC-MS/MS using a metabolic approach. Int J Legal Med 136, 297–308 (2022). https://doi.org/10.1007/s00414-021-02640-w

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