Mineral absorption by submerged bone in marine environments as a potential PMSI indicator

Human remains enter marine environments in a number of ways ranging from homicides, suicides, accidental drownings, shipwrecks, to burials at sea. Once the remains are discovered, a legal and forensic investigation begins. A key component to this investigation is the postmortem submergence interval (PMSI). Determining this range on skeletonized remains is a complicated process in which there is no accurate test; although barnacle growth data was previously used to determine PMSI, there are still limitations with that method. Therefore, a more reliable component of bone needs to be used as a potential PMSI indicator, such as its elemental composition. Diagenesis starts affecting bones immediately and continues for thousands of years. Although diagenesis is a slow process, an exchange of elements between bone and the marine environment continually occurs. The purpose of the present study is to determine whether an increase in marine elements is found within the composition of bone after being submerged in a marine environment for up to 20 months. The present study will also determine whether bones submerged in different aquatic environments have significantly different elemental concentrations. For the time trials, pig femora were submerged in lobster cages off the coast of the University of Massachusetts Boston for 2-20 months. For the salinity trials, pig femora were submerged in a freshwater pond (Holliston, MA), the Inner Boston Harbor, and an ocean inlet near Woods Hole, MA for 18 months. All bone samples were dried, milled, homogenized, and analyzed by ED-XRF under He purge. The initially produced mass percentages of the identified elements were corrected with certified values of standard reference materials (NIST 1486, 1646a, and 2702). A Pearson’s correlation test determined that the concentrations for K, Fe, Zn, Sr, Si, S, Cr, Mn, Cl, Br, Ta, and W were significantly correlated to the amount of time submerged in the water. An ANCOVA analysis was applied to the significant elements noted above. After adjusting for the amount of time submerged, the concentrations of K, Fe, Sr, Si, S, Cl, Br, and Ta were determined to be significantly different between the control samples (never submerged) and the submerged samples (submerged for 2-20 months). K was the only element that had greater concentrations in the control samples than the submerged samples, most likely because of the decrease in mass percent as other environmental elements were incorporated into the bone. S and W were significantly related to the number of months submerged, with S being positively influenced and W being negatively. A multivariable linear regression was run in order to identify a means of predicting the amount of time submerged from the elemental concentrations of an unknown bone from a marine environment. The regression produced an equation that used the concentrations for K, Sr, Si, S, Cr, Cl, and Br to predict the PMSI in months. For the salinity trials, a one-way ANOVA was performed on all the elemental concentrations from the different salinity environments. Post hoc tests determined significant differences in elemental concentrations for K, Fe, Si, S, Al, Ti, Cr, Ni, Mn, Cl and Br among the different submergence locations; elemental concentrations of S, Fe, Mn, Cl, K, and Br were either significantly different between the fresh, brackish, and saltwaters or the freshwater and some form of marine water (brackish and salt). The trends in the other elemental concentrations were less obvious due to the impact of pollution within the surrounding environments. The linear regression equation created in the present study accounted for the majority of the variance in the outcome (R2 = 80.2%); however, this equation should not currently be applied in forensic investigations. The study needs to be repeated a number of times with other bone samples from the same and different submergence locations, in order to determine the accuracy and usefulness of the equation. Although not verified, this regression equation may be useful in analyzing samples from brackish and saltwater environments, because the majority of the variables within the equation (K, Sr, S, Cl, Br) were consistent among the fresh, brackish, and saltwater samples. Time constraints, small sample sizes, and variance among samples were the major limitations of the present study. Even with limitations, significant results were produced by the ED-XRF analysis. Future research should expand upon the methodologies of XRF analyses of bones, especially those from marine environments. Because of their relevance to forensic investigations and PMSI, future research should include longer experimental periods, more salinity locations, more information on the surrounding water components, and more comparisons among instrumentation