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High variation in hypoxanthine determination after analytical treatment of vitreous humor samples

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Abstract

In the field of legal medicine the correct determination of the time of death is of great importance because an error in calculating the post-mortem interval (PMI) could be crucial in a criminal investigation. The quantification of hypoxanthine (Hx) concentration in the vitreous humor (VH) as a means to estimate PMI is useful when dealing with a recent time interval of death and has several advantages over other biological matrices. However, due to its viscous nature, VH has to be liquefied prior to its analysis, and the different procedures to liquefy it may cause alterations in Hx concentration, and hence in estimation of PMI. The aim of this study was to determine differences in Hx concentration after applying several pre-analytical treatments to different aliquots of the same VH sample. Enzymatic digestion, sonication, centrifugation and heat were the liquefying methods applied to reduce the viscosity of 105 VH samples. Analysis was performed using solid phase extraction and liquid chromatography in tandem with mass spectrometry. Statistically significant differences in the measured Hx concentrations were found between enzymatic digestion and the other three methods analyzed (sonication, centrifugation, and heat). Additionally, high data dispersion was found under heat treatment but was not observed for centrifugation or sonication, where similar and more moderate results were found. We conclude that high variations in Hx determination are observed when enzymatic digestion or heat treatment is used and we therefore recommend sonication or centrifugation to quantify Hx concentration in the VH.

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Authors and Affiliations

  1. Institute of Forensic Sciences, University of Santiago de Compostela, 15782, Santiago de Compostela, Spain

    Ana Camba, Elena Lendoiro, María S. Rodríguez-Calvo & José I. Muñoz-Barús

  2. Centre Branch, National Institute of Legal Medicine and Forensic Sciences of Portugal, Coimbra, Portugal

    Cristina Cordeiro & Duarte N. Vieira

  3. Centre of Forensic Sciences (CENCIFOR), University of Coimbra, Coimbra, Portugal

    Cristina Cordeiro & Duarte N. Vieira

  4. Faculty of Medicine, University of Coimbra, Coimbra, Portugal

    Cristina Cordeiro & Duarte N. Vieira

  5. Department of Statistics and O. R., University of Vigo, Vigo, Spain

    Isabel Martínez-Silva

  6. Department of Pathology and Forensic Sciences, University of Santiago de Compostela, Santiago de Compostela, Spain

    María S. Rodríguez-Calvo & José I. Muñoz-Barús

  7. Faculty of Health Sciences, University of Beira Interior, Covilhã, Portugal

    Duarte N. Vieira

Authors
  1. Ana Camba
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  2. Elena Lendoiro
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  3. Cristina Cordeiro
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  4. Isabel Martínez-Silva
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  5. María S. Rodríguez-Calvo
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  6. Duarte N. Vieira
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  7. José I. Muñoz-Barús
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Correspondence to José I. Muñoz-Barús.

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Camba, A., Lendoiro, E., Cordeiro, C. et al. High variation in hypoxanthine determination after analytical treatment of vitreous humor samples. Forensic Sci Med Pathol 10, 627–633 (2014). https://doi.org/10.1007/s12024-014-9590-3

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  • DOI: https://doi.org/10.1007/s12024-014-9590-3

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