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BACKGROUND: The nitrogen isotope ratio (NIR) is a promising index of traditional food intake for an Alaska Native (Yup'ik) population, which can be measured in blood and hair. However, the NIR has not been calibrated to high-quality measures of Yup'ik traditional food intake. OBJECTIVES: Our primary objective was to examine associations between intakes of Yup'ik traditional food groups, including fish, marine mammals, birds, land mammals, berries, greens, and total traditional foods, and the NIR. In an exploratory analysis, we also examined whether NIR analyzed sequentially along hair could reflect dietary seasonality. METHODS: We recruited 68 participants from 2 Yup'ik communities in the Yukon Kuskokwim region of Southwest Alaska (49% female, aged 14–79 y). Participants completed 4 unscheduled 24-h food recalls over the period peak of RBC and hair synthesis preceding a specimen collection visit. The NIR was measured in RBCs ( n = 68), a proximal hair section (n = 58), and sequential segments of hair from individuals in the upper 2 quartiles of traditional food intake having hair >6 cm in length, plus 2 low subsistence participants for reference (n = 18). Diet–biomarker associations were assessed using Pearson's correlation and linear regression. RESULTS: Intakes of fish, marine mammals, berries, and greens were significantly associated with the NIR. The strongest dietary association was with total traditional food intake (R(2) = 0.62), which indicated that each 1‰ increase in the RBC NIR corresponded to 8% of energy from traditional foods. Hair NIR appeared to fluctuate seasonally in some individuals, peaking in the summertime. CONCLUSIONS: Findings support the use of the RBC and hair NIR to assess total traditional food intake in a Yup'ik population. Analyses of sequential hair NIR provided evidence of seasonality in traditional food intake, although seasonal variations were modest relative to interindividual variation.

It is generally believed that early agriculture on the Korean Peninsula was established during the Mumun period (1500–100 BC). While previous studies on agriculture in prehistoric Korea have relied on cultivated plant remains from archaeological sites, only a few isotopic studies have been conducted on Mumun individuals due to poor bone preservation during this period. Here, we measured bulk carbon (δ13C) and nitrogen (δ15N) stable isotope ratios as well as individual amino acid δ13C results (δ13CAA) of collagen from human (n = 7) and animal (n = 4) bones from three Mumun sites (Hwangsok-ri, Jungdo, Maedun Cave) in the central inland portion of South Korea. The aims of this study were to explore the contribution of plant foods to the human diet and to examine the type and extent of agriculture in the Mumun period. In contrast to the surrounding C3 vegetation, all the Mumun humans in this study had significantly 13C-enriched results, evidence for the consumption of C4 plants (foxtail and broomcorn millet). The δ13CAA data show that there was no consumption of freshwater or marine resources in the diet of the Mumun. These data indicate that C4 plants (millets) were the main dietary sources in central inland South Korea and that millet agriculture was fully established during the Middle Mumun period. This finding highlights the importance of millet cultivation during the Mumun period and provides a revaluation for the significance of millets in the development of early agriculture on the Korean Peninsula.

Current approaches to reconstruct subsistence and dietary trends in ancient hunter-gatherer societies include stable isotope analyses, but these have focused on human remains, cooking pottery, and food residues, which are relatively rare in the archaeological record. In contrast, short-term hearths are more ubiquitous worldwide, and these features can provide valuable evidence for ancient subsistence practices, particularly when faunal remains are not preserved. To test the suitability of hearths for this purpose, we conducted multiple chemical analyses: stable carbon and nitrogen isotope analyses of total organic matter (expressed as δ(13)C and δ(15)N values) and compound-specific carbon isotope analyses of individual fatty acids (δ(13)C16:0 and δ(13)C18:0) from 17 well-preserved hearths present in three occupations dating between ∼13,200-11,500 calibrated years B.P. at the Upward Sun River (USR) site in central Alaska. We combined δ(15)N and δ(13)CFA data in a Bayesian mixing model (stable isotope analysis in R) with concentration dependency to each hearth. Our model values were tested against faunal indices, indicating a strong positive relationship between marine proportional contributions to each hearth and salmon abundance. Results of the models show substantial anadromous salmon use in multiple USR components, indicating recurrent use of the site for salmon processing during the terminal Pleistocene. Our results demonstrate that salmonid and freshwater resources were more important for late Pleistocene hunter-gatherers than previously thought and highlight the potential of chemical profiling of hearth organic residues for providing greater geographic and temporal insights into resource use by prepottery societies.

Relict woolly mammoth (Mammuthus primigenius) populations survived on several small Beringian islands for thousands of years after mainland populations went extinct. Here we present multiproxy paleoenvironmental records to investigate the timing, causes, and consequences of mammoth disappearance from St. Paul Island, Alaska. Five independent indicators of extinction show that mammoths survived on St. Paul until 5,600 ± 100 y ago. Vegetation composition remained stable during the extinction window, and there is no evidence of human presence on the island before 1787 CE, suggesting that these factors were not extinction drivers. Instead, the extinction coincided with declining freshwater resources and drier climates between 7,850 and 5,600 y ago, as inferred from sedimentary magnetic susceptibility, oxygen isotopes, and diatom and cladoceran assemblages in a sediment core from a freshwater lake on the island, and stable nitrogen isotopes from mammoth remains. Contrary to other extinction models for the St. Paul mammoth population, this evidence indicates that this mammoth population died out because of the synergistic effects of shrinking island area and freshwater scarcity caused by rising sea levels and regional climate change. Degradation of water quality by intensified mammoth activity around the lake likely exacerbated the situation. The St. Paul mammoth demise is now one of the best-dated prehistoric extinctions, highlighting freshwater limitation as an overlooked extinction driver and underscoring the vulnerability of small island populations to environmental change, even in the absence of human influence.

Palaeoenvironmental records from the now-submerged Bering Land Bridge (BLB) covering the Last Glacial Maximum (LGM) to the present are needed to document changing environments and connections with the dispersal of humans into North America. Moreover, terrestrially based records of environmental changes are needed in close proximity to the re-establishment of circulation between Pacific and Atlantic Oceans following the end of the last glaciation to test palaeo-climate models for the high latitudes. We present the first terrestrial temperature and hydrologic reconstructions from the LGM to the present from the BLB’s south-central margin. We find that the timing of the earliest unequivocal human dispersals into Alaska, based on archaeological evidence, corresponds with a shift to warmer/wetter conditions on the BLB between 14 700 and 13 500 years ago associated with the early Bølling/Allerød interstadial (BA). These environmental changes could have provided the impetus for eastward human dispersal at that time, from Western or central Beringia after a protracted human population standstill. Our data indicate substantial climate-induced environmental changes on the BLB since the LGM, which would potentially have had significant influences on megafaunal and human biogeography in the region. © 2018 The Authors.

This study reports the results of stable isotope analyses (carbon, nitrogen and sulphur) of human bone collagen from the Sunhung mural tomb from the Three Kingdoms Period of Korea (AD 300–668). The stable isotope data indicate that the main source of protein in the diet of the interred seven Sunhung individuals came from C3-based terrestrial resources, and there was an isotopic variation between individuals at this site. To investigate dietary patterns in the Three Kingdoms period, we compared our results with reported isotopic data from other southeastern Three Kingdoms sites (Imdang-dong, Songhyeon-dong and Yean-ri). We found that the Sunhung individuals had similar isotope ratios to the people from these other sites. However, there was noticeable isotopic difference among individuals from each study site, although much of the dietary protein in each site was mainly from terrestrial sources. We propose that the most parsimonious explanation for this isotopic pattern is variation in social status during this time period. Copyright © 2012 John Wiley & Sons, Ltd.

We analyzed stable carbon and nitrogen isotope ratios of human and animal remains from the Ando shell midden, South Korea. The Ando site is a rare Incipient Chulmun (Neolithic) site (ca. 6000–5000 BC), which contains well-preserved human and animal bones in shell mounds. The stable isotope results for humans (average δ 13 C = −13.5 ± 0.5‰ and δ 15 N = 15.2 ± 0.5‰) indicate that Ando people in the Incipient Chulmun period strongly depended on marine resources. There were no isotopic differences between humans of different sex and age at this site. We compared our data with other previous published isotopic data from the Chulmun sites and found that the Ando people had similar isotope values to the southern Chulmun people (Tongsamdong and Daepo), but different isotopic ratios than the western Chulmun people (Daejukri and Konamri). These results indicate that marine foods were the main food resources in the southern coastal regions, but not in the western coastal regions in Chulmun Korea.

This research presents the individual amino acid δ13C values in bone collagen of humans (n = 9) and animals (n = 27) from two prehistoric shell midden sites in Korea. We obtained complete baseline separation of 16 of the 18 amino acids found in bone collagen by using liquid chromatography–isotope ratio mass spectrometry (LC–IRMS). The isotopic results reveal that the humans and animals in the two sites had similar patterns in essential amino acids (EAAs) and non-essential amino acids (NEAAs). The EAA and NEAA δ13C values in humans are intermediate between those in marine and terrestrial animals. However, the threonine δ13C values in humans and animals measured in this study are more highly enriched than those of other amino acids. At both sites, all amino acids in marine animals are 13C-enriched relative to those of the terrestrial animals. The isotopic evidence suggests that the Tongsamdong human had EAAs and NEAAs from marine food resources, while the Nukdo humans mainly had EAAs from terrestrial food resources but obtained NEAAs from both terrestrial and marine resources. The δ13C isotopic differences in amino acids between marine and terrestrial animals were the largest for glycine (NEAA) and histidine (EAA) and the smallest for tyrosine (NEAA) and phenylalanine (EAA). In addition, threonine among the EAAs also had a large difference (∼8‰) in δ13C values between marine and terrestrial animals, and has the potential to be used as an isotopic marker in palaeodietary studies. Threonine δ13C values were used in conjunction with the established Δ13CGlycine–phenylalanine values and produced three distinct dietary groups (terrestrial, omnivorous, and marine). In addition, threonine δ13C values and Δ13CSerine–phenylalanine values were discovered to separate between two dietary groups (terrestrial vs. marine), and these δ13C values may provide a potential new indicator for investigating the distinction between marine and terrestrial protein sources in human diets.

This article presents the first isotopic investigation of human and animal bone remains from the Middle Chulmun (3500–2000 BC) period in southeastern Korea. We have obtained a single human and associated faunal stable carbon and nitrogen isotope results from the Tongsamdong site, a coastal shell midden. Despite the discovery of domesticated plants and the existence of large amounts of terrestrial mammal bones from the shell midden, the human and dogs we measured were heavily dependent on marine protein resources for their lives. Although our stable isotope results are based on a small number of individuals due to the lack of human remains at this period, isotopic evidence suggests the possibility that Tongsamdong people in the Middle Chulmun period depended largely on marine protein resources. This isotopic evidence is consistent with the archaeological evidence from the site.

Carbon and nitrogen stable isotope analysis was undertaken on bone collagen extracted from archaeological human ( n = 48) and animal ( n = 45) skeletons from the Nukdo site, Location I C, South Korea. This shell midden and grave site is dated from the late Mumun (550–300 BC) to early Iron Age (300 BC-1 AD) periods. The herbivorous mammals fell within the range of C 3 consumers, with average values of δ 13 C = −21.0 ± 0.5‰ and δ 15 N = 3.6 ± 0.5‰ for deer ( n = 16) and δ 13 C = −20.6 ± 0.5‰ and δ 15 N = 4.5 ± 2.0‰ for wild boar ( n = 17). Humans from this site averaged δ 13 C = −18.3 ± 0.4‰ and δ 15 N = 11.2 ± 0.7‰ for adults ( n = 15) and δ 13 C = −18.7 ± 0.7‰ and δ 15 N = 12.5 ± 1.1‰ for juveniles ( n = 33). These δ 13 C values indicate that there was no significant input of C 4 plants in the human diets and this may be associated with the spread of rice agriculture in the Mumun period. Human bone collagen δ 13 C and δ 15 N values indicate that there was some consumption of marine foods, although the main protein sources were from terrestrial foods. The isotope data demonstrate that the humans at Nukdo had mixed diets that included marine and terrestrial protein, including C 3 plants such as rice. Finally, the isotope results from the juveniles indicate that weaning occurred before the age of 1.5 years in this period.

We report a three-phase chromatographic method for the separation and analysis of delta(13)C values of underivatized amino acids from biological proteins (keratin, collagen, and casein) using liquid chromatography-isotope ratio mass spectrometry (LC-IRMS). Both precision and accuracy of delta(13)C values for standard amino acid mixtures over the range of approximately 8 to 1320 ng of carbon per amino acid on the column were assessed. The precision of delta(13)C values of amino acids was found to be better at higher concentrations, whereas accuracy improved at lower concentrations. The optimal performance for this method was achieved with between 80 and 660 ng of carbon of each amino acid on the column. At amino acid amounts lower than 20 ng of carbon on the column, precision and accuracy may become compromised. The application of this new three-phase chromatographic technique will allow the analysis of delta(13)C of amino acids to be carried out as a routine method and benefit fields of research such as biomedicine, forensics, ecology, nutrition, and palaeodiet reconstruction in archaeology.

An objective dietary biomarker would help clarify the contribution of sugar-sweetened beverage (SSB) intake to obesity and chronic disease risk. Previous studies have proposed the carbon isotope ratio (δ(13)C) as a biomarker of SSB intake but found associations that were of modest size and confounded by other components of the diet. We investigated whether the δ(13)C values of nonessential amino acids (δ(13)CNEAA) in RBCs could provide valid biomarkers that are more specific to SSBs. We assessed the associations of RBC δ(13)CNEAA with SSB intake in a study population of 68 Yup'ik people, using gas chromatography/combustion/isotope ratio mass spectrometry to measure δ(13)CNEAA and four 24-h dietary recalls to assess intake. Among RBC nonessential amino acids, alanine δ(13)C (δ(13)Calanine) was strongly correlated with intake of SSBs, added sugar, and total sugar (r = 0.70, 0.59, and 0.57, respectively; P < 0.0001) but uncorrelated with other dietary sources of elevated δ(13)C. We also evaluated whether sweetener intake could be noninvasively assessed using hair δ(13)Calanine in a subset of the study population (n = 30). Hair δ(13)Calanine was correlated with RBC δ(13)Calanine (r = 0.65; P < 0.0001) and showed similar associations with SSB intake. These results show that δ(13)Calanine in RBCs provides a valid and specific biomarker of SSB intake for the Yup'ik population and suggest RBCs and hair δ(13)Calanine as candidate biomarkers of SSB intake for validation in the general U.S. population. Ultimately, these biomarkers could clarify our understanding of whether and how SSB intake contributes to chronic disease.

Stable carbon and nitrogen isotope analyses were conducted to investigate dietary variation in human skeletons (n = 109) from the Gaya cemetery at Yeanri located near Gimhae City, South Korea. The cemetery contained three distinct grave types dating to 4th-7th century AD. The main purposes of this research were to reconstruct palaeodiet in the Gaya population and to explore correlations between stable isotope compositions and burial types, inferred age, and sex of these individuals. The isotopic data indicate that the people at Yeanri consumed a predominantly C(3)-based terrestrial diet supplemented with freshwater and/or marine resources. The comparison of isotopic results reveals significant differences in delta(13)C values among three adult burial types (wood-cist coffin: -18.5 +/- 0.5 per thousand, stone-cist coffin: -18.1 +/- 0.6 per thousand, mausoleum: -17.8 +/- 0.9 per thousand). Males in wood-cist and stone-cist coffins have relatively more elevated mean delta(13)C and delta(15)N values than females. The isotopic ratios from the two adult age groups (21-40 years and 40-60 years) indicate that there was no significant dietary change in individuals with age. The isotope data from the infants and children suggest the weaning was a gradual process that was completed between 3 and 4 years of age in the Gaya population. This evidence indicates that the dietary variations within the cemetery reflect social status, sex, and childhood consumption patterns.

Supplementary methods and results

Supplementary methods and results