Analysis of biogenic amines in vitreous humor using high performance liquid chromatography for the estimation of postmortem interval
- WCU Author/Contributor (non-WCU co-authors, if there are any, appear on document)
- Savannah Reyh Mayer (Creator)
- Institution
- Western Carolina University (WCU )
- Web Site: http://library.wcu.edu/
- Advisor
- Nuwan Perera
Abstract: A novel method is proposed to estimate the postmortem interval (PMI) using the biogenic amine concentration in vitreous humor. There are multiple methods that can be used to determine the time of death (PMI). However, these methods often have uncertainties that complicate estimating an accurate PMI. To address this challenge and improve the accuracy in PMI determination, this study focuses on utilizing biogenic amine levels in vitreous humor. Biogenic amines, formed from the decarboxylation of proteins during anaerobic bacterial degradation following discontinuation of oxygen transport at death, exhibit increased concentrations in body tissues and surrounding areas. Vitreous humor is a valuable specimen collected during the autopsies for the analysis of various drugs and metabolites. It is an anatomically isolated specimen and crucial information on the presence of drugs such as alcohol and other substances at the time of death due to the lack of enzymes. The formation of biogenic amines in vitreous humor may occur at a slower rate compared to other tissues in the body and thus this matrix can provide more accurate information on PMI. A high-performance liquid chromatographic (HPLC) method was developed to analyze biogenic amines in vitreous humor as a part of this thesis research. Baseline separation of amines of interest including methylamine, cadaverine, putrescine, histamine, tyramine, and heptylamine was achieved. For a more accurate representation of the biogenic amine concentrations 1,7-diaminoheptane was used as an internal standard to adjust for any matrix effects. Porcine eyes were chosen to be representative of human eyes due to the similarities in vitreous humor volume and composition. The eyes were let to decompose under three different environmental conditions to evaluate the impact environmental conditions have on the PMI. One interset of eyes was placed in polyethylene bags after the collection and let to decompose at room temperature. The second set of eyes were frozen for three days before the decomposition in polyethylene bags. The third set of eyes were placed in a large polyethylene box and let to decompose at room temperature. In these three experiments analysis indicated that methylamine, putrescine cadaverine, and histamine are present in the vitreous humor. Concentrations of methylamine, putrescine, cadaverine, and histamine are best represented in the frozen eye experiment to estimate PMI. Additionally, eyes then were put directly into the sealed bag had predictable concentration patterns of methylamine, putrescine, and cadaverine. The results from the third experiment (boxed eyes) were inconclusive as they did not have any predictable concentration correlation over time particularly for cadaverine and histamine, therefore, should not be used for PMI estimation.
Analysis of biogenic amines in vitreous humor using high performance liquid chromatography for the estimation of postmortem interval
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Created on 4/1/2024
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Additional Information
- Publication
- Thesis
- Language: English
- Date: 2024
- Keywords
- Biogenic Amines, HPLC, Post Mortem Interva, Vitreous Humor
- Subjects
- Biogenic amines
- High performance liquid chromatography
- Death—Time of
- Vitreous humor