The Age Determination of Body Fluids Using Alternative Forensic Techniques Charlotte-Maria Orphanou BSc (Hons), Dr. Harry Mountain and Dr. Laura Walton-Williams c.orphanou@staffs.ac.uk The Forensic Science Society & California Association of Criminalists joint Autumn Conference Forensic Horizons 2013 Version 0.2

INTRODUCTION The most commonly encountered body fluids at crimes scenes: Blood Semen Saliva Vaginal secretions Age determination is the next significant step for forensic analysis in criminal investigations.

RELEVANCE Police: Place an individual at the scene of a crime, before, during, or after. Support or refute an alibi. Provide intelligence information that could link scenes and people of interest.

RELEVANCE Scenes of Crime Officers: Any biological evidence can be collected using the normal procedures. No additional risk of contamination or misinterpretation of pre-treatment measures.

RELEVANCE Forensic Practitioners: Age determination analysis could provide a timeline of events. Current accepted method is expensive and time consuming. Establishing a method that could be routinely used by forensic practitioners.

MY RESEARCH Blood, saliva and vaginal secretions aged to determined time intervals. Alternative forensic techniques: ATR-FTIR: examines reflectance of component vibrations within the infrared region. SDS PAGE: separates proteins by molecular weight.

SAMPLE COLLECTION & STORAGE Deposited onto 100% cotton cloth. Stored in accordance with the Human Tissue Act (2004). Aged to predetermined time intervals ranging from 24 hours to 18 months.

FTIR PRELIMINARY RESEARCH Explored blood, saliva and vaginal secretions. Showed potential in a small aging study from 24 hours to 7 days. Demonstrated that identification of body fluid components was needed.

Figure 1: Neat wet sample spectra a) Blood b) Saliva c) Vaginal Secretions Figure 1: Neat wet sample spectra a) blood, b) saliva, c) vaginal secretions.

Figure 2: Neat air dried sample spectra after 5 hours a) Blood b) Saliva c) Vaginal Secretions Figure 2: Neat air dried sample spectra after 5 hours a) blood, b) saliva, c) vaginal secretions.

SDS-PAGE PRELIMINARY RESEARCH Method development: Extraction Running Conditions Identifying protein components. Measuring protein degradation over time. a) Saliva b) Blood Figure 3: SDS-PAGE protein separation a) saliva, b) blood.

RESULTS TO DATE FTIR demonstrates blood peak intensity variation over time. a) Blood Figure 4: a) Blood overlay spectra of 24 hours (purple), 14 days (green) and 21 days (red).

RESULTS TO DATE Saliva and vaginal secretions undetectable on cotton when using FTIR. a) Saliva b) Vaginal secretions Figure 5: a) Saliva overlay spectra of neat 5 hours air dried (red), 7 days (turquoise) & cotton blank (bright green); b) Vaginal secretions overlay spectra of 5 hours air dried (red), 7 days (orange) & cotton blank (green).

ONGOING RESEARCH SDS-PAGE method development and application to aged samples. Storage of long term aging samples (18 months) is underway. FTIR instrument comparison to ensure reproducibility amongst differing instruments.

FURTHER RESEARCH Investigating the age determination of semen. Deposition of body fluids on different materials. Effect of freezing storage on age determination.

USEFUL REFERENCES Elkins, K.M. (2011) Journal of Forensic Sciences, 56 (6) 1580-1587. Chiriboga, L. et al. (1998) Biospectroscopy, 4 47-53. D’Amici, G.M. Rinalducci, S. and Zolla, L. (2007) Journal of Proteome Research, 6 3242-3255. Garidel, P. and Schott, H. (2006) BioProcess International, 40-46. Garidel, P. and Schott, H. (2006) BioProcess International, 48-55. Lamy, et al. (2012) Intech, DOI: 10.1007/s00414-013-0848-1 Movasaghi, Z. Rehman, S. and ur Rehman, I. (2008) Applied Spectroscopy Reviews 43, 134-179. Scott, D.A. et al. (2010) Diabetology and Metabolic Syndrome, 2 (48). Yang, H. et al. (2013) International Journal of Legal Medicine, DOI: 10.1007/s00414-013-0848-1

ACKNOWLEDGEMENTS The Forensic Science Society Staffordshire University

Thank you for listening Questions? c.orphanou@staffs.ac.uk