Disaster victim identification: Stable isotope analysis and the identification of unknown decedents.

Within the complex world of disaster victim identification, or DVI, forensic science practitioners use a variety of investigative techniques to work toward a common goal: identification of the decedents, bringing closure to the affected communities. Identification is a complex undertaking; the event (disaster) also can be extraordinarily complex, as it may be an acute event, or one that spans months or years. Compounding this time issue, remains may be heavily fragmented, dispersed, commingled, or otherwise disrupted by either the perpetrators or the disaster itself. To help solve these complexities, we explore the use of stable isotope analysis (SIA) in DVI events. SIA can be used with a variety of body tissues (hair, nail, bone, and teeth), and each represents different time depths in a decedent's life. Bone collagen and tooth enamel carbonate are useful to reconstruct an individual's diet and source water intakes, respectively, leading to likely population or geographic origin determinations. Additionally, the carbon and nitrogen isotopic signatures of bone collagen have calculated intraperson ranges. These facts allow investigators to determine likely origin of remains using isotopic data and can be used to link skeletal elements (to an individual), or perhaps more importantly, show that remains are not linked. Application of SIA can thus speed remains identification by eliminating individuals from short lists for identification, linking or decoupling remains, and reducing the need for some DNA testing. These strategies and hypothesis tests should commence early in the DVI process to achieve maximum effectiveness.

Forensic investigation of falsified antimalarials using isotope ratio mass spectrometry: a pilot investigation.

We explored whether isotope ratio mass spectrometry (IRMS) is useful to investigate the origin of falsified antimalarials. Forty-four falsified and genuine antimalarial samples (artesunate, artemether-lumefantrine, dihydroartemisinin-piperaquine and sulphamethopyrazine-pyrimethamine) were analyzed in bulk for carbon (C), nitrogen (N), and oxygen (O) element concentrations and stable isotope ratios. The insoluble fraction ("starch") was extracted from 26 samples and analyzed. Samples of known geographical origin maize, a common source of excipient starch, were used to produce a comparison dataset to predict starch source. In both an initial (n = 18) and a follow-on set of samples that contained/claimed to contain artesunate/artemether (n = 26), falsified antimalarials had a range of C concentrations less than genuine comparator antimalarials and δ13C values higher than genuine comparators. The δ13C values of falsified antimalarials suggested that C4 plant-based organic material (e.g., starch derived from maize) had been included. Using the known-origin maize samples, predictions for growth water δ18O values for the extracted "starch" ranged from - 6.10 to - 1.62‰. These findings suggest that IRMS may be a useful tool for profiling falsified antimalarials. We found that C4 ingredients were exclusively used in falsified antimalarials versus genuine antimalarials, and that it may be possible to predict potential growth water δ18O values for the starch present in falsified antimalarials.

Forensic application of isotope ratio mass spectrometry (IRMS) for human identification.

Application of isotope ratio mass spectrometry (IRMS) to skeletal remains has become an important tool to investigate human behavior and history. Isotopic variations in collagen, enamel, and keratin reflect variations in an individual's diet and drinking water. Since food and water sources typically are geographically linked, isotope testing can assist in forensic identification by classifying remains to a likely geographic or population origin. If remains are commingled, differences in diet or geographic origin also can support their separation. The usefulness of IRMS in forensic science is dependent on the underlying quality and surety of the isotope test results; in other words, we need to understand their reliability in interpretations. To take ownership of isotopic data quality, we recommend asking a series of questions:Here, we use data collected during the buildout and accreditation of an isotope testing program at the Defense POW/MIA Accounting Agency (DPAA) to answer the above questions for the forensic application of IRMS for human identification. While our primary focus is on the preparation and analysis of bone collagen, the questions above should be considered whenever isotope testing is used in forensic casework. Whether the populations of interest are drugs or humans, olives or explosives, users need to evaluate their isotopic data and interpretations to ensure they are scientifically sound and legally defensible.

A large-scale evaluation of intraperson isotopic variation within human bone collagen and bioapatite.

This study investigated intraperson skeletal (herein referred to as either "intraperson" or "intraskeletal") variation in stable isotope ratios for collagen (C and N) and bioapatite (C and O) extracted from five to six long bones from 27 modern individuals. The maximum intraperson variation observed for collagen was 0.78‰ for δ13Ccoll values and 1.12‰ for δ15Ncoll values, with a mean variation (± SD) of 0.33 ± 0.18‰ and 0.45 ± 0.27‰, respectively. For bioapatite, the maximum intraperson variation was 1.63‰ for δ13Cap values and 4.80‰ for δ18Oap values, with a mean variation (± SD) of 0.81 ± 0.32‰ and 1.00 ± 1.03‰, respectively. These results generally agree with previously reported data on intraskeletal isotopic variation. Using a two- and three-standard-deviations-from-the-mean model with analytical quality control data included, it is proposed that two bones with differing collagen δ13Ccoll values greater than 0.75‰ are probably from different individuals, and those that have differing values greater than 0.95‰ are from different individuals. Likewise, differing collagen δ15Ncoll values greater than 1.05‰ are probably different, and greater than 1.35‰ are different. For bioapatite, the proposed values change to 1.55‰ and 1.90‰ for δ13Cap, respectively; for δ18Oap values no limits were set due to the unexpectedly large variation found in the study population. We highly encourage researchers to use extreme caution when interpreting δ18O values from bone apatite. We also note that these parameters were evaluated on modern samples and therefore may not reflect intraperson variation in past societies. Finally, we demonstrate application of these interpretative limits to sort commingled human remains cases.

Recent applications of isotope analysis to forensic anthropology.

Isotope analysis has become an increasingly valuable tool in forensic anthropology casework over the past decade. Modern-day isotopic investigations on human remains have integrated the use of multi-isotope profiles (e.g. C, N, O, H, S, Sr, and Pb) as well as isotopic landscapes ("isoscapes") from multiple body tissues (e.g. teeth, bone, hair, and nails) to predict possible region-of-origin of unidentified human remains. Together, data from various isotope analyses provide additional lines of evidence for human identification, including a decedent's possible region-of-birth, long-term adult residence, recent travel history, and dietary choices. Here, we present the basic principles of isotope analysis and provide a brief overview of instrumentation, analytical standards, sample selection, and sample quality measures. Finally, we present case studies that reflect the diverse applications of isotope analysis to the medicolegal system before describing some future research directions. As shown herein, isotope analysis is a flexible and powerful geolocation tool that can provide new investigative leads for unidentified human remains cases.

Lessons learned from inter-laboratory studies of carbon isotope analysis of honey.

Forensic application of carbon isotope ratio measurements of honey and honey protein to investigate the degree of adulteration with high fructose corn syrup or other C4 plant sugars is well established. These measurements must use methods that exhibit suitable performance criteria, particularly with regard to measurement uncertainty and traceability - low levels of adulteration can only be detected by methods that result in suitably small measurement uncertainties such that differences of 1‰ or less can be reliably detected. Inter-laboratory exercises are invaluable to assess the state-of-the art of measurement capabilities of laboratories necessary to achieve such performance criteria. National and designated metrology institutes from a number of countries recently participated in an inter-laboratory assessment (CCQM-K140) of stable carbon isotope ratio determination of bulk honey. The same sample material was distributed to a number of forensic isotope analysis laboratories that could not participate directly in the metrological comparison. The results from these studies have demonstrated that the majority of participants provided isotope delta values with acceptable performance metrics; that all participants ensured traceability of their results; and that where measurement uncertainties were reported; these were fit-for-purpose. A number of the forensic laboratories only reported precision rather than full estimates of measurement uncertainty and this was the major cause of the few instances of questionable performance metrics. Reporting of standard deviations in place of measurement uncertainties is common practice outside metrology institutes and the implications for interpretations of small differences in isotopic compositions are discussed. The results have also highlighted a number of considerations that are useful for organisers of similar inter-laboratory studies in the future.

Strontium isotope ratios of human hair from the United States: Patterns and aberrations.

RATIONALE: Strontium isotope ratios (87 Sr/86 Sr) of hair may be a valuable tool to estimate human provenance. However, the systematics and mechanisms controlling spatial variation in 87 Sr/86 Sr of modern human hair remain unclear. Here, we measure 87 Sr/86 Sr of hair specimens from across the USA to assess the presence of geospatial relationships. METHODS: Ninety-eight human hair specimens were collected from salon/barbershop floors in 48 municipalities throughout the conterminous USA. [Sr] and 87 Sr/86 Sr ratios were measured from hair using quadrupole and multi-collector inductively coupled plasma mass spectrometers, respectively. The [Sr] and 87 Sr/86 Sr ratios of hair were compared with the measured [Sr] and 87 Sr/86 Sr ratios of tap waters from the collection locations. In addition, the 87 Sr/86 Sr ratio of hair was compared with the modeled ratios of bedrock and surface waters. RESULTS: Hair color was independent of the 87 Sr/86 Sr ratio, but related to [Sr]. The 87 Sr/86 Sr ratios of hair and leachate were not statistically different and were positively correlated; however, in several hair-leachate pairs, the ratios were conspicuously different. The 87 Sr/86 Sr ratios of both hair and leachate were linearly correlated with tap water. The 87 Sr/86 Sr ratio of hair was also significantly correlated with the modeled ratio of bedrock and surface waters, although the 87 Sr/86 Sr ratio of hair was most strongly correlated with the measured ratio of tap water. CONCLUSIONS: The 87 Sr/86 Sr ratio of hair is related to the ratio of tap water, which varied geographically. The ratio of hair provided geographic information about an individual's recent residence. Differences in the 87 Sr/86 Sr ratios of hair and hair leachate may be concomitant with travel and could potentially be used as a screening tool to identify recent movements.

Applying the principles of isotope analysis in plant and animal ecology to forensic science in the Americas.

The heart of forensic science is application of the scientific method and analytical approaches to answer questions central to solving a crime: Who, What, When, Where, and How. Forensic practitioners use fundamentals of chemistry and physics to examine evidence and infer its origin. In this regard, ecological researchers have had a significant impact on forensic science through the development and application of a specialized measurement technique-isotope analysis-for examining evidence. Here, we review the utility of isotope analysis in forensic settings from an ecological perspective, concentrating on work from the Americas completed within the last three decades. Our primary focus is on combining plant and animal physiological models with isotope analyses for source inference. Examples of the forensic application of isotopes-including stable isotopes, radiogenic isotopes, and radioisotopes-span from cotton used in counterfeit bills to anthrax shipped through the U.S. Postal Service and from beer adulterated with cheap adjuncts to human remains discovered in shallow graves. Recent methodological developments and the generation of isotope landscapes, or isoscapes, for data interpretation promise that isotope analysis will be a useful tool in ecological and forensic studies for decades to come.

TATP isotope ratios as influenced by worldwide acetone variation.

Isotope ratio analysis has been shown to discriminate samples of forensic interest and to link many synthesized and natural materials to their precursors when traditional chemical and physical analyses cannot. Successful application of stable isotope analysis to chemicals of interest requires a background of likely variations in stable isotope ratios; often, this background population can be generated from analysis of possible precursors and the relationships of stable isotopes of precursor(s) to product(s), which may depend on synthesis techniques. Here we measured the carbon (13C/12C) and hydrogen (2H/1H) isotope ratios of the oft-illicitly manufactured explosive triacetone triperoxide, TATP, and one of its precursors, acetone. As acetone is the sole source of carbon and hydrogen to TATP, a survey of acetone from 12 countries was conducted to explore the breadth of 13C/12C and 2H/1H variation in the precursor, and therefore, its product. Carbon and hydrogen isotope ratios were measured using continuous flow isotope ratio mass spectrometry (IRMS) techniques. We observed greater ranges in both C and H isotope ratios of acetone than previously published; we also found that country-of-purchase was a large contributing factor to the observed variation, larger than acetone grade and brand. Following clandestine production methods, we observed that the stable isotope ratios of TATP retained the stable isotope signatures of acetone used in synthesis. We confirmed the robustness of TATP carbon isotope ratios to both recrystallization and time-dependent sublimation, important considerations when faced with the task of practical sampling of potential unexploded TATP from a crime scene.

Stable hydrogen and oxygen isotopes of tap water reveal structure of the San Francisco Bay Area's water system and adjustments during a major drought.

Water availability and sustainability in the Western United States is a major flashpoint among expanding communities, growing industries, and productive agricultural lands. This issue came to a head in 2015 in the State of California, when the State mandated a 25% reduction in urban water use following a multi-year drought that significantly depleted water resources. Water demands and challenges in supplying water are only expected to intensify as climate perturbations, such as the 2012-2015 California Drought, become more common. As a consequence, there is an increased need to understand linkages between urban centers, water transport and usage, and the impacts of climate change on water resources. To assess if stable hydrogen and oxygen isotope ratios could increase the understanding of these relationships within a megalopolis in the Western United States, we collected and analyzed 723 tap waters across the San Francisco Bay Area during seven collection campaigns spanning 21 months during 2013-2015. The San Francisco Bay Area was selected as it has well-characterized water management strategies and the 2012-2105 California Drought dramatically affected its water resources. Consistent with known water management strategies and previously collected isotope data, we found large spatiotemporal variations in the δ2H and δ18O values of tap waters within the Bay Area. This is indicative of complex water transport systems and varying municipality-scale management decisions. We observed δ2H and δ18O values of tap water consistent with waters originating from snowmelt from the Sierra Nevada Mountains, local precipitation, ground water, and partially evaporated reservoir sources. A cluster analysis of the isotope data collected in this study grouped waters from 43 static sampling sites that were associated with specific water utility providers within the San Francisco Bay Area and known management practices. Various management responses to the drought, such as source switching, bringing in new sources, and water conservation, were observed in the isotope data. Finally, we estimated evaporative loss from one utility's reservoir system during the 2015 water year using a modified Craig-Gordon model to estimate the consequences of the drought on this resource. We estimated that upwards of 6.6% of the water in this reservoir system was lost to evaporation.

Reconstruction of travel history using coupled δ18 O and 87 Sr/86 Sr measurements of hair.

RATIONALE: Oxygen isotope ratios (δ18 O values) of hair largely reflect features of regional hydrology while strontium isotope ratios (87 Sr/86 Sr) are thought to reflect bedrock geology; combination of both isotope signatures may provide greater capacity for determining provenance and reconstructing travel history of an organism. To test this hypothesis, we compared the O-Sr isotope profiles of hair from domestic horses with known residency histories. METHODS: Tail hairs were collected from a pair of horses pastured together for a period of 16 months, one of which lived in a different location for the 8 months prior. Hair samples were washed with solvents to remove external contaminants prior to sequential sampling for δ18 O and 87 Sr/86 Sr analysis via TC/EA-IRMS and MC-ICP-MS, respectively. Hair digests were concentrated and analyzed employing low-flow natural aspiration to measure 87 Sr/86 Sr. RESULTS: Tail hair from the control and transported horses had mean δ18 O values of 11.25 ± 1.62 ‰ and 10.96 ± 1.53 ‰, and mean 87 Sr/86 Sr of 0.7101 ± 0.0006 and 0.7109 ± 0.0020, respectively. The δ18 O and 87 Sr/86 Sr profiles for the control and transported horses were indistinguishable when they were pastured together. The 87 Sr/86 Sr profiles were significantly different during the period that the horses were living apart, while the δ18 O values were indistinguishable during that period. CONCLUSIONS: By comparing the O-Sr isotope profiles of a control and transported horse, we investigated isotopic signal(s) potentially useful for reconstructing travel histories via high-resolution sequential sampling along single strands of tail hair. Improved analytical capabilities allowed for extremely low Sr abundance samples to be analyzed for 87 Sr/86 Sr and proved capable of resolving a horse's movement between distinct regions. Copyright © 2017 John Wiley & Sons, Ltd.

Radiocarbon dating of seized ivory confirms rapid decline in African elephant populations and provides insight into illegal trade.

Carbon-14 measurements on 231 elephant ivory specimens from 14 large ivory seizures (≥0.5 ton) made between 2002 and 2014 show that most ivory (ca 90%) was derived from animals that had died less than 3 y before ivory was confiscated. This indicates that the assumption of recent elephant death for mortality estimates of African elephants is correct: Very little "old" ivory is included in large ivory shipments from Africa. We found only one specimen of the 231 analyzed to have a lag time longer than 6 y. Patterns of trade differ by regions: East African ivory, based on genetic assignments of geographic origin, has a much higher fraction of "rapid" transit than ivory originating in the Tridom region of Cameroon-Gabon-Congo. Carbon-14 is an important tool in understanding patterns of movement of illegal wildlife products.

The influences of cultivation setting on inflorescence lipid distributions, concentrations, and carbon isotope ratios of Cannabis sp.

While much is known about how the growth environment influences many aspects of floral morphology and physiology, little is known about how the growth setting influences floral lipid composition. We explored variations in paraffin wax composition in Cannabis sp., a cash crop grown both indoors and outdoors across the United States today. Given an increased focus on regulation of this crop, there are additional incentives to certify the setting of Cannabis cultivation. To understand the impacts of the growth environment, we studied distributions, concentrations, and carbon isotope ratios of n-alkanes isolated from Cannabis sp. inflorescences to assess if variations within these lipid parameters were related to known growth settings of specimens seized by federal agents. We found that Cannabis plants cultivated under open-field settings had increased inflorescence paraffin wax abundances and greater production of lower molecular weight n-alkanes relative to plants grown in enclosed environments. Further, the carbon isotope ratios of n-C29 from Cannabis plants grown in enclosed environments had relatively lower carbon isotope (δ(13)C) values compared to plants from open-field environments. While this set of observations on seized plant specimens cannot address the particular driver behind these observations, we posit that (a) variations in irradiance and/or photoperiod may influence the distribution and concentration of inflorescence lipids, and (b) the δ(13)C value of source CO2 and lipid concentration regulates the δ(13)C values of inflorescence n-C29 and bulk Cannabis plant materials. Nonetheless, by using a cultivation model based on δ(13)C values of n-C29, the model correctly identified the growth environment 90% of time. We suggest that these lipid markers may be used to trace cultivation methods of Cannabis sp. now and become a more powerful marker in the future, once the mechanism(s) behind these patterns is uncovered.

Improved accuracy and precision in δ15 NAIR measurements of explosives, urea, and inorganic nitrates by elemental analyzer/isotope ratio mass spectrometry using thermal decomposition.

RATIONALE: Elemental analyzer systems generate N(2) and CO(2) for elemental composition and isotope ratio measurements. As quantitative conversion of nitrogen in some materials (i.e., nitrate salts and nitro-organic compounds) is difficult, this study tests a recently published method - thermal decomposition without the addition of O(2) - for the analysis of these materials. METHODS: Elemental analyzer/isotope ratio mass spectrometry (EA/IRMS) was used to compare the traditional combustion method (CM) and the thermal decomposition method (TDM), where additional O(2) is eliminated from the reaction. The comparisons used organic and inorganic materials with oxidized and/or reduced nitrogen and included ureas, nitrate salts, ammonium sulfate, nitro esters, and nitramines. Previous TDM applications were limited to nitrate salts and ammonium sulfate. The measurement precision and accuracy were compared to determine the effectiveness of converting materials containing different fractions of oxidized nitrogen into N(2). RESULTS: The δ(13) C(VPDB) values were not meaningfully different when measured via CM or TDM, allowing for the analysis of multiple elements in one sample. For materials containing oxidized nitrogen, (15) N measurements made using thermal decomposition were more precise than those made using combustion. The precision was similar between the methods for materials containing reduced nitrogen. The %N values were closer to theoretical when measured by TDM than by CM. The δ(15) N(AIR) values of purchased nitrate salts and ureas were nearer to the known values when analyzed using thermal decomposition than using combustion. CONCLUSIONS: The thermal decomposition method addresses insufficient recovery of nitrogen during elemental analysis in a variety of organic and inorganic materials. Its implementation requires relatively few changes to the elemental analyzer. Using TDM, it is possible to directly calibrate certain organic materials to international nitrate isotope reference materials without off-line preparation.

The potential for application of ink stable isotope analysis in questioned document examination.

We investigated a novel application of stable isotope abundance analysis of nitrogen (15N), carbon (13C), hydrogen (2H), and oxygen (18O) to characterize pen ink. We focused on both ballpoint and gel pen inks. We found that the isotope ratios of ink from pens purchased together in a package were similar and within-package stable isotope ratio variability was not significantly larger than the variability of isotope reference materials used during analysis. In contrast, the isotope ratios of ink from pens of the same brand purchased in three states of the continental USA were significantly different from each other and there was isotope ratio variation among pens of the same brand but different, unknown production periods. The stable isotope ratios of inked paper were statistically distinguishable using measured δ15N values. Paper inked with different gel pens was statistically distinguishable using measured δ2H values. The capacity of stable isotope ratios to differentiate among ballpoint inks as well as gel inks shows that stable isotope analysis may be a useful and quantifiable investigative technique for questioned document examination, although current sample size requirements limit its utility. Application of the technique in casework will require the development of micro-scale sampling and analysis methods.

Deuterium content of water increases depression susceptibility: the potential role of a serotonin-related mechanism.

Environmental factors can significantly affect disease prevalence, including neuropsychiatric disorders such as depression. The ratio of deuterium to protium in water shows substantial geographical variation, which could affect disease susceptibility. Thus the link between deuterium content of water and depression was investigated, both epidemiologically, and in a mouse model of chronic mild stress. We performed a correlation analysis between deuterium content of tap water and rates of depression in regions of the USA. Next, we used a 10-day chronic stress paradigm to test whether 2-week deuterium-depleted water treatment (91 ppm) affects depressive-like behavior and hippocampal SERT. The effect of deuterium-depletion on sleep electrophysiology was also evaluated in naïve mice. There was a geographic correlation between a content of deuterium and the prevalence of depression across the USA. In the chronic stress model, depressive-like features were reduced in mice fed with deuterium-depleted water, and SERT expression was decreased in mice treated with deuterium-treated water compared with regular water. Five days of predator stress also suppressed proliferation in the dentate gyrus; this effect was attenuated in mice fed with deuterium-depleted water. Finally, in naïve mice, deuterium-depleted water treatment increased EEG indices of wakefulness, and decreased duration of REM sleep, phenomena that have been shown to result from the administration of selective serotonin reuptake inhibitors (SSRI). Our data suggest that the deuterium content of water may influence the incidence of affective disorder-related pathophysiology and major depression, which might be mediated by the serotoninergic mechanisms.

Carbon and nitrogen isotope ratios of factory-produced RDX and HMX.

RDX and HMX are explosive compounds commonly used by the military and also occasionally associated with acts of terrorism. The isotopic characterization of an explosive can be a powerful approach to link evidence to an event or an explosives cache. We sampled explosive products and their reactants from commercial RDX manufacturers that used the direct nitration and/or the Bachmann synthesis process, and then analyzed these materials for carbon and nitrogen isotope ratios. For manufacturers using the Bachmann process, RDX (13)C enrichment relative to the hexamine substrate was small (+0.9‰) compared to RDX produced using the direct nitration process (+8.2‰ to +12.0‰). RDX (15)N depletion relative to the nitrogen-containing substrates (-3.6‰) was smaller in the Bachmann process than in the direct nitration process (-12.6‰ to -10.6‰). The sign and scale of these differences agree with theorized mechanisms of mass-dependent fractionation. We also examined the isotopic relationship between RDX and HMX isolated from explosive samples. The δ(13)C and δ(15)N values of RDX generally matched those of the HMX with few exceptions, most notably from a manufacturer known to make RDX using two different synthesis processes. The range in δ(13)C values of RDX in a survey of 100 samples from 12 manufacturers spanned 33‰ while the range spanned by δ(15)N values was 26‰; these ranges were much greater than any previously published observations. Understanding the relationship between products and reactants further explains the observed variation in industrially manufactured RDX and can be used as a diagnostic tool to analyze explosives found at a crime scene.

Deconvolution of isotope signals from bundles of multiple hairs.

Segmental analysis of hair has been used in diverse fields ranging from forensics to ecology to measure the concentration of substances such as drugs and isotopes. Multiple hairs are typically combined into a bundle for segmental analysis to obtain a high-resolution series of measurements. Individual hair strands cycle through multiple phases of growth and grow at different rates when in the growth phase. Variation in growth of hair strands in a bundle can cause misalignment of substance concentration between hairs, attenuating the primary body signal. We developed a mathematical model based on the known physiology of hair growth to describe the signal averaging caused by bundling multiple hairs for segmental analysis. The model was used to form an inverse method to estimate the primary body signal from measurements of a hair bundle. The inverse method was applied to a previously described stable oxygen isotope chronology from the hair of a murder victim and provides a refined interpretation of the data. Aspects of the reconstruction were confirmed when the victim was later identified.

Isolation of strontium pools and isotope ratios in modern human hair.

The elements of human hair record specific information about an individual's health, diet, and surrounding environment. Strontium isotope ratios of human hair have attracted interest as they potentially record an individual's environment. Yet, separating the external environmental signals from the internal dietary indicators has remained a challenge. Here, we examined the effects of five different hair-cleaning methodologies to determine the extent that internal and external strontium signals can be isolated from human hair. In the first study of its kind, we employed an in-line strontium purification methodology and a multi-collector inductively coupled plasma mass spectrometer to obtain high-precision strontium isotope ratio of human hair and of leachates of the different washing treatments. We found that the different applications of an individual treatment removed a consistent amount of strontium from hair and that replicate analyses showed each treatment altered the strontium isotope ratios of hair consistently. A mass-balance approach was applied to demonstrate that strontium was quantitatively removed and was accounted for in either the treated hair or the leachate. We observed that strontium isotope ratio varied as a function of treatment aggressiveness so as to suggest that there was a fine-scale structuring of strontium within hair (transverse cross-sectional variations); these variations existed as differences in strontium concentrations and isotope ratios. As a result, the Sr isotope ratio of hair and hair leachates treated with the most aggressive cleaning methods reflected the isotope ratios of the interior and total exterior strontium signatures, respectively. The results of this study indicate that external environmental strontium signals can be distinguished from the internal signals and therefore permit the application of strontium isotope ratios of modern human hair for geospatial applications.

Dietary heterogeneity among Western industrialized countries reflected in the stable isotope ratios of human hair.

Although the globalization of food production is often assumed to result in a homogenization of consumption patterns with a convergence towards a Western style diet, the resources used to make global food products may still be locally produced (glocalization). Stable isotope ratios of human hair can quantify the extent to which residents of industrialized nations have converged on a standardized diet or whether there is persistent heterogeneity and glocalization among countries as a result of different dietary patterns and the use of local food products. Here we report isotopic differences among carbon, nitrogen and sulfur isotope ratios of human hair collected in thirteen Western European countries and in the USA. European hair samples had significantly lower δ(13)C values (-22.7 to -18.3‰), and significantly higher δ(15)N (7.8 to 10.3‰) and δ(34)S (4.8 to 8.3‰) values than samples from the USA (δ(13)C: -21.9 to -15.0‰, δ(15)N: 6.7 to 9.9‰, δ(34)S: -1.2 to 9.9‰). Within Europe, we detected differences in hair δ(13)C and δ(34)S values among countries and covariation of isotope ratios with latitude and longitude. This geographic structuring of isotopic data suggests heterogeneity in the food resources used by citizens of industrialized nations and supports the presence of different dietary patterns within Western Europe despite globalization trends. Here we showed the potential of stable isotope analysis as a population-wide tool for dietary screening, particularly as a complement of dietary surveys, that can provide additional information on assimilated macronutrients and independent verification of data obtained by those self-reporting instruments.