Investigating the presence of ancient absorbed organic residues in groundstone using GC–MS and other analytical techniques: a residue study of several prehistoric milling tools from central California

Analysis of absorbed organic molecules in groundstone could provide a valuable means to study resource use and processing in antiquity. The following study analyzed extracts from the surfaces of several central California milling tools to assess whether organic residues remained from prehistoric resource processing. It also sought to determine which source identification methods are likely to be successful at providing information about the type, or even the specific identify, of resources that were processed. Lipids (primarily fatty acids) were analyzed using GC–MS and the presence of phenolic compounds was assessed with UV-Vis spectroscopy. Milling surfaces were compared to previously broken surfaces from the same tool with the assumption that both surfaces had been exposed to similar post-depositional conditions. Results supported the presence of ancient residues in milling tools. A higher concentration of fatty acids was recovered from milling surfaces than paired broken surfaces. Furthermore, measurable amounts of azelaic acid (an oxidation product of some unsaturated fatty acids) were present in most milling surfaces, but not in broken surfaces. However, results also indicated that environmentally absorbed lipids formed a significant portion of the total lipid content. Thus, it is suggested that future analyses employ a biomarker approach, rather than criteria based on ratios of common fatty acids, to identify sources of organic residues in prehistoric milling tools.     

The meals in a Tuscan building yard during the Middle Age. Characterization of organic residues in ceramic potsherds

The characterization of organic residues found in ceramics objects coming from the Carmine Convent in Siena (Central Italy), probably built in the second half of the 13th century, is described. The nine objects analyzed in this study are pots, casseroles, and lids and they were used as filling material of the vault of one room of the convent.     

Chemical analyses of organic residues in archaeological pottery from Arbon Bleiche 3, Switzerland – evidence for dairying in the late Neolithic

Fatty acids distribution and stable isotope ratios (bulk δ¹³C, δ¹⁵N and δ¹³C of individual fatty acids) of organic residues from 30 potsherds have been used to get further insights into the diet at the Late Neolithic (3384–3370 BC) site of Arbon Bleiche 3, Switzerland. The results are compared with modern equivalents of animal and vegetable fats, which may have been consumed in a mixed ecology community having agrarian, breeding, shepherd, gathering, hunting, and fishing activities. The used combined chemical and isotopic approach provides valuable information to complement archaeological indirect evidence about the dietary trends obtained from the analysis of faunal and plant remains. The small variations of the δ¹³C and δ¹⁵N values within the range expected for degraded animal and plant tissues, is consistent with the archaeological evidence of animals, whose subsistence was mainly based on C₃ plants. The overall fatty acid composition and the stable carbon isotopic compositions of palmitic, stearic and oleic acids of the organic residues indicate that the studied Arbon Bleiche 3 sherds contain fat residues of plant and animal origin, most likely ruminant (bovine and ovine). In several vessels the presence of milk residues provides direct evidence for dairying during the late Neolithic in central Europe.     

蜂蜡类残留物的科学分析研究

蜂蜡是古代人类可获得的最常见的天然材料之一，在人类生产生活中占有重要地位。对蜂蜡类残留物的科技分析可直接提供古代人类对蜂蜡利用的科学证据。本文介绍了蜂蜡的产生与组成，梳理了蜂蜡类残留物的样品处理与检测分析方法，并总结了蜂蜡类残留物科学鉴定的依据，以期为古代蜂蜡利用与其他相关问题的研究提供有益参考。     

Swept under the rug: the problem of unacknowledged ambiguity in lithic residue identification

Microscopic analysis of organic residues on stone tools is used to interpret prehistoric stone tool functions. The morphology of some residues can be difficult to interpret, yet this ambiguity is rarely acknowledged in the literature. Our research seeks to understand the nature of this ambiguity by objectively identifying ambiguous residues in our reference collection. We trained four archaeologists in residue analysis using one part of our reference collection, then tested their ability to identify sixty-eight residues in another part of the same collection. Forty-eight of the residues in the test (70%) were correctly identified by three or all four subjects. We considered the remaining twenty residues, which were correctly identified by two or fewer of the subjects, to be ambiguous. These are most often in the hide-scraping, bone-scraping, and hardwood-scraping (macerated) categories, and tend to have an atypical morphology which falls in the range of variability of another residue category. Some of these residues also have optical properties which make them more difficult to image than others. We explore the potential for scanning electron microscopy (SEM) to improve residue identification in a second test. This test shows a modest improvement in identification success rates of ambiguous residues when SEM images are included. We conclude that while images from different types of microscopes can improve reliability of identification, some residues will always be ambiguous. Rather than being ignored, these ambiguities should be brought to light, closely examined, and published as such.     

An Optimized Approach for Protein Residue Extraction and Identification from Ceramics After Cooking

Extraction of protein residues from archaeological matrices, such as pottery clay, lithics, and grinding stones, has proven to be methodologically challenging. Protein residue analysis is fraught with technical challenges in analytical chemistry. In cooking pottery, protein residues are thought to bind to clay surfaces in vessel walls through a variety of primarily non-covalent interactions. Removal of protein residues requires the disruption of these interactions, and a diverse set of tools has been proposed and applied. Here, we test extraction procedures through varying combinations of physical parameters and solvents to derive an optimal approach yielding efficiencies of recovery from experimental pottery above 60%. The utility of our extraction approach was further validated through liquid chromatography?Cmass spectrometry analysis of experimental residues. We have identified several hurdles to developing a successful study of protein residues from pottery, each of which is surmountable with additional method development in the realm of archaeological chemistry.     

Remains of the day-preservation of organic micro-residues on stone tools

Here I report on the decay processes of microscopic organic residues left on stone tool surfaces after their use. Residue analysis on ancient stone tools facilitates reconstruction of past activities. This study enables predictions about the circumstances under which ancient residues preserve. Experimental tool sets with modern residues were buried for a year in separate deposits at Sterkfontein, Sibudu (South Africa) and Zelhem (the Netherlands) whose pH and geomorphology varied, they were then analysed using light microscopy. Biological weathering mainly causes residue decay. In unstable environments rich in microbes and micro-organisms, residues decay quickly. From an archaeological perspective this means that sites that are stable, desiccated, waterlogged, extremely acidic or alkaline and extremely cold or hot sites. Different residue types have different preservation optima and this may lead to a preservation and perhaps interpretation bias. The preliminary predictive models presented in this paper could aid in the considered selection of sites and samples.     

Small things in perspective: the contribution of our blind tests to micro-residue studies on archaeological stone tools

Our blind tests are distinctive for they were conducted on replicated stone tools used for a variety of tasks that included the processing of animal remains and plants. The analyst was required to differentiate an array of residues from microscopic morphological characteristics, using light microscopy. The original aim of our first tests was to assess the analyst's ability to identify a variety of plant and animal residues, but issues and problems that arose during the testing process made it clear that greater value might be gained from the lessons that we learnt about methodology and the direction for future micro-residue research. We show that problems identified during our first tests stimulated research. Amongst other things, we learnt to distinguish plant and animal remains more confidently than previously. Our residue analyses are firmly embedded in wider archaeological research and our tests help to explain why there are sometimes contradictions between the evidence from archaeologically recovered remains and residues on stone tools. A further outcome of the tests is that we have adopted a multi-stranded approach that provides a cautious, but secure strategy for identifying and interpreting use-residues. Our studies of contaminants through replications have also been invaluable for distinguishing incidental residues from use-related residues.     

Recovery of Protein and DNA Trapped in Stone Tool Microcracks

Microcracks produced during the manufacture of stone tools may harbour ancient biological residues. Our studies test this hypothesis using light, scanning electron, fluorescence, and confocal microscopy to characterize microcracks produced in obsidian microblades by pressure flaking. Cell-sized fluorescent latex beads penetrated microcracks to depths exceeding 50 μm below the microblade surface. Fluorescently labelled blood protein and DNA were trapped in microcracks within 5 min of exposure. Washing artifacts with water did not remove blood residues from microcracks. However, 60–80% of trapped blood protein and DNA were released by an 18-hour incubation in 4M guanidine hydrochloride or by sonication in 5% ammonium hydroxide. These experiments suggest that residues trapped in artifact microcracks represent an important and often overlooked source of ancient DNA and protein.     

Late Pleistocene to Early Holocene Plant Movements in Southern Kyushu, Japan

From the late Pleistocene to early Holocene in Japan, subtropical and temperate forest elements moved northwards. This affected human choices and access to food sources. More settled patterns of living spread northwards gradually, and northern hunting-gathering-fishing people began cultivating vegetables and cereal crops. This poster reports the presence of ancient starch residues on stone artefacts in Kagoshima Prefecture, southern Kyushu. The oldest residues recovered are dated by context to about 30,000 ¹⁴C yr BP. If such residues can be identified, it may be possible to detect a hypothesized early phase of tropical plant movement northwards during warmer climate peaks in the late terminal Pleistocene, as well as during the long period of Holocene warming that followed. As an initial step towards identification, the morphological characteristics and condition of the starch granules are described and compared to those of other sites in early Japan.     

Potential misidentification of in situ archaeological tool-residues: starch and conidia

Microscopic identification of organic residues in situ on the surface of archaeological artefacts is an established procedure. Where soil components morphologically similar to use-residue types exist within the soil, however, there remains the possibility that these components may be misidentified as authentic residues. The present study investigates common soil components known as conidia, fungal spores which may be mistaken for starch grains. Conidia may exhibit the rotating extinction cross under cross-polarised light commonly diagnostic of starch, and may be morphologically indistinguishable from small starch grains, particularly at the limits of microscope resolution. Conidia were observed on stone and ceramic archaeological artefacts from Honduras, Palau and New Caledonia, as well as experimental artefacts from Papua New Guinea. The findings act as a caution that in situ analysis of residues, and especially of those less than 5 μm in size, may be subject to misidentification.     

Starch residues on museum artefacts: implications for determining tool use

A sample of 11 museum artefacts was examined to assess the preservation of any adhering organic residues, particularly starch granules, and to assess the potential for the identification of cooked starchy foods. The primary aim was to examine starchy deposits, extract starch granules and examine their physical condition to increase our understanding of the taphonomy of starch granules and associated organic residues in archived material and archaeological deposits. The study shows that organic plant residues could be recovered from archived artefacts and makes a significant contribution to understanding the function of this material. Recovered starch granules showed an interesting pattern of varying organic preservation on the surface of stone and wooden artefacts. Some starch granules were found to be well preserved while others in the same sample appeared to have been affected by enzyme degrading micro-organisms. Recovered residues included fully gelatinised starch.     

Chemical Residues as Anthropic Activity Markers. Ethnoarchaeology,Experimental Archaeology and Archaeology of Food Production and Consumption

ABSTRACT

Chemical residues preserved in floors can be considered anthropic activity markers. In fact, residues are strictly related to the activities performed and reflect their spatial distribution. We present a synthesis of the work carried out over the last few decades in Mexico and Italy related to the study of chemical residues in floors. Residues can be identified performing specific chemical analyses both of plastered or earthen floors samples. We outline a methodological approach concerning the use of the markers of the activities to interpret food production and consumption in the archaeological record, based upon evidence from experimental, ethnoarchaeological and archaeological examples. Here we point out the advantages and problems of such an approach, mainly related to equifinality, of the use of spot tests and gas chromatography coupled with mass spectrometry techniques using examples from different sites in the world.     

Combining residue analysis of floors and ceramics for the study of activity areas at the Garum Shop at Pompeii

In this paper, we propose the application—for the first time in the Mediterranean area—of the combination of the study of chemical residues in floors and ceramics, with the aim of providing information about the activities carried out in archeological buildings. We chose the Garum Shop at Pompeii to test the method. In fact, due to the peculiarity of this archeological context, it provided an ideal case in which the activities performed are in part known, and the ceramic vessels recovered are still in situ. Floor samples were studied by means of spot tests developed in Mexico aimed at identifying the presence of phosphates, fatty acids, and protein residues, while the organic residues preserved in the ceramic matrix of amphorae, dolia, and other ceramic vessels were studied by gas chromatography coupled with mass spectrometry. Moreover, we integrated the data obtained with specific studies directed at better identifying the solid residues found inside two of the amphorae studied: botanical studies of fruit stones recovered in a Dressel 20 amphora and the characterization of the lime preserved in an African amphora. The research allowed for the identification of the traces of some of the activities performed, such as cooking and producing garum in the floors of the building, and the use and re-use of amphorae and dolia before the Vesuvian eruption.     

The use of an improved pRIA technique in the identification of protein residues

Blood and protein residue identification in archaeological research has been a controversial subject for the last 20 years. This paper reports on the use of an improved protein radioimmunoassay (pRIA) technique in identifying protein residues. Results from the blind testing of the original and improved pRIA techniques in identifying bloodstains on experimental lithic artifacts are described and compared. We argue that the improved pRIA technique is highly sensitive and accurate for identifying protein residues to genus, and thus has excellent applications for archaeological research.     

Social Spaces of Daily Life: A Reflexive Approach to the Analysis of Chemical Residues by Multivariate Spatial Analysis

Studying human activities requires an examination of the inherent epistemological problems in building arguments about the past based on chemical residues and modern observations. A reflexive approach to the analysis of chemical residues at the San Lucas archaeological site, a Classic Hohokam settlement located in Marana, Arizona, represents a unique opportunity to evaluate current techniques and paradigms for the interpretation of daily life activities. By incorporating an innovative program rooted in satellite remote sensing image analysis and spatial statistics, including new techniques, such as bulk density, loss on ignition, electrical conductivity, and salinity, results suggest that soil chemical analysis will benefit more from learning about structure and agency than from one single activity.     

Clues to Stone Tool Function Re-examined: Comparing Starch Grain Frequencies on Used and Unused Obsidian Artefacts

The identification of plant residues observed on prehistoric stone artefacts has often been used to inform on tool function without adequate consideration of potential contamination due to post-depositional processes. The paper redresses this balance by proposing a methodology which combines use-wear and residue analyses with systematic testing of depositional context to distinguish residues formed during tool use from accidental contamination. A case study involving obsidian artefacts from an open site in Papua New Guinea is used to illustrate the new approach. Starch grains, a class of residue which has had little attention outside Australia, were chosen as the focus of study because of their abundance at this site. The frequency of starch grains extracted from residues on stone artefacts was compared with those in sediments adhering to and adjacent to the artefact. A use-wear study conducted as a blind test provided an independent measure of whether the artefacts had been used. The frequency of starch grains was shown to be significantly correlated with used artefacts and not correlated with unused artefacts or the sediments. The case study demonstrates the importance of testing for contamination and also illustrates the benefit of studying starch grains as a class of plant residue.     

Identifying biomolecular origins of solid organic residues preserved in Iron Age Pottery using DTMS and MVA

The chemical characterisation of solid organic residues found in ceramic vessels, can provide archaeologists with valuable information about ancient diets. Although several selective analytical techniques have been applied to the analysis of specific classes of compounds, such as extractable lipids, waxes, terpenoids and protein fragments, a non-selective analytical technique is required to characterise and categorise complete solid organic residues. In this study, Direct Temperature-resolved Mass Spectrometry (DTMS) is used for the characterisation of 34 solid residues situated on vessels recovered from an indigenous settlement from the Roman period at Uitgeest – Groot Dorregeest (The Netherlands). Sample preparation is limited to grinding very small samples (5–10 μg) and suspending them in water. DTMS analysis of aliquots (1–2 μL) of this suspension gave information about a broad range of organic compounds, such as lipids, polynuclear aromatic hydrocarbons, markers for residual proteins and polysaccharides, and for newly formed complex condensed polymers. Multivariate analysis of the DTMS spectra identified five different chemotypes: groups of residues with comparable chemical characteristics. The biomolecular origin of each of these chemotypes is identified by comparison with experimentally charred reference materials. The chemotypes A₁ and A₂ consist of charred residues identified as starch-rich foods (mixed with animal or plant products), chemotype C consists of protein-rich charred animal products without starch, chemotype B contains smoke condensates from wood fires, and chemotype D consists of special protein-rich and lipid-free foods or non-food products.     

Extracting residues from stone tools for optical analysis: towards an experiment-based protocol

The identification of residues is traditionally based on the distinctive morphologies of the residue fragments by means of light microscopy. Most residue fragments are amorphous, in the sense that they lack distinguishing shapes or easily visible structures under reflected light microscopy. Amorphous residues can only be identified by using transmitted light microscopy, which requires the extraction of residues from the tool’s surface. Residues are usually extracted with a pipette or an ultrasonic bath in combination with distilled water. However, a number of researchers avoid residue extraction because it is unclear whether current extraction techniques are representative for the use-related residue that adheres to a flaked stone tool. In this paper, we aim at resolving these methodological uncertainties by critically evaluating current extraction methodologies. Attention is focused on the variation in residue types, their causes of deposition and their adhesion and on the most successful technique for extracting a range of residue types from the stone tool surface. Based on an experimental reference sample in flint, we argue that a stepwise extraction protocol is most successful in providing representative residue extractions and in preventing damage, destruction or loss of residue.     

Hafted armatures and multi-component tool design at the Micoquian site of Inden-Altdorf,Germany

Excavation of the Micoquian site Inden-Altdorf (Weisweiler-124) near the former German capital Bonn in western Germany has revealed the first valid open-site habitation features with hut-like structures and associated hearths for the Middle Palaeolithic in Central Europe. It has been dated to the Eemian interglacial (OIS 5e), a warm interglacial between 128 and 115 ka BP. Various wear traces and especially organic residues have been detected on a large number of stone tools using microscopic use-wear analysis of lithics recovered from the site. A multi-level analysis developed through an experimental framework and archaeological study using optical light microscopes, scanning electron microscopes and energy-dispersive X-ray microprobes identified the adhering residues as birch pitch. Birch pitch is the oldest synthetically produced material and was used as an adhesive to attach lithic implements to wooden shafts. While such hafting technology is commonly associated with modern humans in the Upper Palaeolithic, the birch pitch residues found on the Micoquian tools of Inden-Altdorf suggest that hafting technologies and the frequent use of multi-component tools already existed in the Middle Palaeolithic, c. 120 ka BP in central Europe.