The practical application of reflectance spectrophotometry for the demonstration of haemoglobin and its degradation in bruises

Abstract

Aims: To develop a non-invasive method to demonstrate the presence of haemoglobin and its degradation products in bruises in live human subjects for the purposes of objectively assisting in the determination of the age of a bruise. Methods: The cuvette holder unit of a Cary 100 Bio UV-Visible Spectrophotometer was replaced with the manufacture's fibre optic cable and optical reflectance probe. The probe was placed on the skin surface. The absorption spectrum from 780 to 380 nm was collected and transformed into the first derivative. Calculation of the first derivative permits absorption attributed to haemoglobin degradation (primarily to bilirubin, but also haemosiderin) to be separated from absorption by haemoglobin. First derivative and colorimetry values, expressed as CIEL*a*b data, were derived from scans of 50 bruises. Results: The fibre optic cable and probe allowed the spectrophotometer to collect reproducible absorption spectra of bruises in the skin of living subjects. A bruise at three days has greater negative first derivative values at 480 and 490 nm than does a fresh bruise, indicating the local degradation of haemoglobin. Correlation between the first derivative and the CIEL*a*b ''b'' values in a series of bruises indicates that the yellow colour in a bruise is proportional to the amount of local haemoglobin breakdown. Conclusion: The ability to demonstrate the presence of haemoglobin and measure its degradation in bruises in living human subjects by a non-invasive method has not been described previously, and may be of use in the objective ageing of bruises for forensic purposes. B y visual inspection it may be possible to suggest that a bruise is either recent (fresh) or old (more than 18 hours), based on the presence or absence of yellow colour.

Skin changes following minor trauma

“…The visual appearance of a bruise will be dependent on the time after impact, and optical methods can be used to monitor and characterize the observed changes. Reflection spectroscopy has been applied to characterize bruises with good results [12][13][14][15]. Bohnert et al [12] published a relation between reflectance spectra, observed bruise color and depth of injury in post mortem cases.…”

“…They found that superficial hemorrhages cause bright red bruises, while deep subcutaneous bruises appear blue or purple. Hughes et al [13] used reflectance spectroscopy to quantify bilirubin in bruises of various age, and Randeberg et al [14] identified hemoglobin content in the skin, hemoglobin oxygenation, and bilirubin content as potential indicators of bruise age. Randeberg et al [15] later published a mathematical model to predict the age of a bruise from a reflectance measurement.…”

Origin and assessment of bruises in beef cattle at slaughter

“…This change in bruise colour is due to the inflammatory Strappini, Metz, Gallo and Kemp process, whereby macrophages are recruited to the injured area and ingest red blood cells and metabolize the haemoglobin first to biliverdin and then rapidly to bilirubin (Hughes et al, 2004). Based on empirical observations, Grandin (2000) concluded that in beef cattle carcasses it would be possible to separate bruises into at least two categories: fresh bruises and bruises that are several days or weeks old.…”

“…The latter accounts for the numerous studies carried out in recent decades with the aim of developing a method for ageing bruises in human skin (Langlois and Gresham, 1991;Betz, 1994;Sawaguchi et al, 2000;Bariciak et al, 2003;Bonelli et al, 2003;Hughes et al, 2004 andRandeberg et al, 2006;Grellner and Madea, 2007;Kondo, 2007).…”

Alternate Light Source Findings of Common Topical Products

“…Depending on its wavelength, light can penetrate the skin and has the potential to detect bruises, which are primarily located below the level of the epidermis (>15μm) within the vascular dermis (Hughes, Ellis, Burt, & Langlois, 2004;Langlois & Gresham, 1991;Sandby-Moller, Poulsen, & Wulf, 2003;Wright & Golden, 2010). At this point, light most commonly interacts with the tissues in three ways: reflection, absorption or fluorescence (Marin & Buszka, 2013).…”