Development of spectroscopic identification and analysis of haemoglobin components in bruises

Author: Josie Nunn

Nunn, Josie, 2020 Development of spectroscopic identification and analysis of haemoglobin components in bruises, Flinders University, College of Science and Engineering

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Bruises form with blunt trauma to the surface of the skin that ruptures vessels, releasing blood which breaks down into biliverdin and bilirubin exhibiting a variety of discolouration over time. Identification of bruise age is crucial in forensic investigations into suspicious deaths, however current methods of gathering information from the breakdown process of blood can be misleading. The original contribution to knowledge in this thesis is the development of spectroscopic (particularly far infrared) identification of differences in key indicators of a bruise's age; haemoglobin breakdown components.

Analysis with an ASD FieldSpecĀ®4 Hi-Res: high resolution spectroradiometer coupled to a 150 mm integrating sphere has potential for mobile and rapid preliminary scanning in the UV-Vis.-NIR region of the electromagnetic spectrum but NIR sensitivity is low. A main concern is water, a physiological solvent, which was found to strongly absorb in the IR region.

MIR and FIR out-of-vacuum ATR analyses at Flinders University and the Australian Synchrotron (THz/FIR beamline) of samples in solid and liquid phase at various concentrations (low to high) presented distinguishable spectra of biliverdin and bilirubin. Different vibrations presented, some attributed to the presence or lack of intramolecular hydrogen bonding in bilirubin, may be fundamental information for identification of early to late stages of bruise breakdown, hence establishment of age.

Additionally, FIR transmission analysis of pure material, PE and KBr pellets provided extended wavenumber range (with Siolo, Si:B and MCTN detectors on the THz beamline) for key biliverdin and bilirubin differences. However, Diamond Window Liquid Cells require further research and development as little to no characteristic vibrations are detected when samples are analysed in dilute liquid phase, possibly due to fundamentally different interactions in liquid phase as compared to solid state.

Keywords: bruise, bruises, blood, haemoglobin, hemoglobin, haemoglobin break down, hemoglobin break down, bilirubin, biliverdin, ferritin, discolouration, bruise age, forensic, pathology, infrared, spectroscopy, haemoglobin breakdown products, asd fieldspec, integrating sphere, ultraviolet, visible spectroscopy, nir, near infrared, mid infrared, far infrared, synchrotron, attenuated total reflectance, australian synchrotron, thz, terrahertz, solid phase, liquid phase, intramolecular hydrogen bonding, pellets, beamline, Josie Nunn, Stewart Walker, Neil Langlois

Subject: Chemistry thesis

Thesis type: Doctor of Philosophy
Completed: 2020
School: College of Science and Engineering
Supervisor: Stewart Walker