Lipid Residues Research Articles

Direct evidence from lipid residue analysis for the routine consumption of millet in Early Medieval Italy

Millets have been cultivated in Europe since the Late Neolithic but, beyond recording their presence, little is known about their use and context of consumption. As a C4 plant, the contribution of millet on diet can be readily identified through stable isotope analysis of human bones. Using this approach, however, previous studies have been unable to distinguish direct consumption of the cereal from the consumption of millet fed animals. Historical evidence suggests that the latter was common practice. To address this issue, we present the first direct evidence for millet consumption in Medieval period using organic residue analysis. Lipid were extracted from 45 pottery vessels from the Episcopal centre in Padua, Northern Italy dating from the 6th to 10th centuries AD. Miliacin, a biomarker for broomcorn millet, was present in many of the cooking vessels tested. Based on the co-occurrence of miliacin with other food derived lipids and the vessel typologies, we suggest that millet was a common culinary ingredient during the Early Medieval period in this region. The earliest evidence dates to the 6th c. AD and notably derives from deposits associated with high status occupation of the site, a surprising result given the common association of these crops as low-status or starvation foods in the historic periods. It is likely that millet was a common cereal staple in human diet during this period in North-eastern Italy and that its use was far less restricted than previously thought. More broadly, our study highlights the efficacy of combining organic residue analysis and stable isotope analysis of bone to relate culinary and dietary information of ancient populations.

Pastoralist Foodways Recorded in Organic Residues from Pottery Vessels of Modern Communities in Samburu, Kenya

Organic residue analyses of archaeological ceramics can provide important insights into ancient foodways. To date, however, there has been little critical reflection on how lipid residues might (or might not) reflect dietary practices or subsistence strategies more generally. A combination of ethnoarchaeological research and chemical and isotopic analyses of lipid residues from pottery made and used by modern Samburu pastoralists in northern Kenya was undertaken to supplement the interpretive framework used in archaeological investigations. A total of 63 potsherds were collected from various contexts, including settlement sites and rockshelters, and analysed using gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS) and gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The results showed that the free fatty acids, palmitic (C16:0) and stearic (C18:0), dominated the lipid profiles, with extremely broad ranges of δ13C values. The majority of the Δ13C values from Samburu pots suggest that vessels were intensively used to process ruminant carcass products, yet the Samburu economy is not, in fact, meat-based at all. Despite an overall reliance on dairy products, milk is rarely processed in ceramic vessels, largely due to cultural prohibitions. Surprisingly, a number of vessels from one site, Naiborkeju Hill, were used to process dairy products. Compound-specific radiocarbon dating of lipids from these sherds suggests that this pottery originated from an earlier period, demonstrating a possible shift in ceramic use by pastoralist communities in this region over time. The overall conclusion is that lipid residues may not necessarily reflect, in a simple way, the day-to-day consumption or the perceived relative importance of different foodstuffs. In the Samburu case, lipid residues reflect the functional and ideological suitability of ceramics for processing only certain types of food (meat/fat/bones), despite an overall reliance on milk. These conclusions are important when considering the origins and development of African pastoralism, for example, as interpreted from the archaeological record.

Lipid Residues from Low‐Fired Pottery

This paper assesses the fate of lipids associated with low‐temperature and pit‐fired pottery to determine to what degree organic compounds persist or are removed during short‐firing episodes below 800 °C. Three different types of clay were fired using contrasting techniques including at 400 °C for 4 h in a muffle furnace, and pit‐firing in which pottery was fired to higher temperatures but for shorter periods of time. Total lipid extracts obtained by solvent extraction of test sherds were screened using gas chromatography‐flame ionization detection (GC‐FID) to determine the lipid concentrations and gas chromatography–mass spectrometry (GC–MS) to identify the organic compounds present. The results showed that firing of clay removes all naturally occurring alkyl lipids; however, during pit‐firing, diterpenoid lipids were introduced into the clay as a condensate from pine ( Pinus spp.) wood used as fuel. These results confirm that alkyl lipids, e.g., fatty acids, can be reliably associated with the use of vessels, although caution is required when interpreting the origins of lipids that might derive from fuel used in firing.

Investigating the function of prehistoric stone bowls and griddle stones in the Aleutian Islands by lipid residue analysis

AbstractThe earliest durable cooking technologies found in Alaska are stone bowls and griddle stones recovered from the Aleutian Islands. This article aims to identify the function of these artefacts. Molecular and chemical analyses of carbonised residues found on their surfaces confirm that these artefacts were used to process marine resources. Both artefacts have high lipid content and C:N ratios, suggesting they were used to process oily substances. Stable isotope results of individual lipids suggest that they were used to process different sets of resources within the aquatic spectrum as griddle stones have slightly more 13C-depleted lipids than stone bowls, possibly indicating more variable use. Integration of these results with archaeological and ethnographic data leads us to infer that griddle stones were used for cooking a diversity of aquatic resources, possibly with the addition of plant foods, whereas stone bowls were specifically used to render marine mammal fats. We further hypothesize that a sudden peak in stone bowl frequencies at 4000–3000 cal yr BP was connected to a Neoglacial cold spell bringing sea ice conditions to the Aleutian Islands. This may have led to new subsistence strategies in which the rendering of marine mammal fats played a central role.

Integration of Linearbandkeramik cattle husbandry in the forested landscape of the mid-Holocene climate optimum: Seasonal-scale investigations in Bohemia

Domestic animals and plants were introduced to Europe from the Near East and subsequently spread across Europe, entailing adaptations to different environments with consequences for the biology of organisms, agropastoral technical systems and socio-economic organisation. Agriculture was introduced to Central Europe by Linearbandkeramik (LBK) societies between 5600 and 4900 cal. BC, in predominantly forested environments. LBK farming systems involved intensive permanent field cultivation in natural openings. Milking was practiced as evidenced from cattle mortality profiles and lipid residues in ceramics. Questions arise as to what extent LBK cattle husbandry relied on woodland, and as to whether the seasonal scarcity of fodder conditioned cattle reproduction cycles, with consequences on milk availability. Results from the δ13C and δ18O analysis of cattle tooth enamel at Chotěbudice and Černý Vůl (Bohemia, Czech Republic) suggest a limited use of dense forest for cattle herding, even on a seasonal scale: cattle were kept in the open component of the forest/steppe mosaic landscape. Winter forest browsing/provision of leafy fodder was evidenced in one specimen. At Chotěbudice, cattle births mainly occurred over a two to three-month period, suggesting environmental constraints on cattle fertility cycles, and possibly seasonal fodder scarcity. A direct consequence of this would be a shorter period of milk availability throughout the year.

Towards the detection of dietary cereal processing through absorbed lipid biomarkers in archaeological pottery

The uptake of cereal agriculture in the Neolithic is one of the most important processes in later human prehistory. However, in many parts of Europe, early evidence from pollen or macrofossils is scarce or inconclusive, and there are considerable ambiguities about timing, intensity and the mode of transition to agriculture in these regions.An alternative approach is organic residue analysis, a technique that targets lipids preserved in the walls of unglazed ceramic pots used for storage and processing of foodstuffs. By analysing the molecular and isotopic composition of absorbed lipid residues, many different food items and processing techniques can be detected and distinguished. However, this approach is by-and-large limited to animal-based food sources, and despite their importance, many plant-based food items including cereals are currently not accessible with this approach.For a better understanding of the behaviour of cereal lipids, cooking experiments were conducted and the uptake of cereal-specific compounds such as alkylresorcinols and plant sterols into the ceramic matrix was investigated using a new sensitive method based on GC-Q-ToF-MS. Furthermore, changes in the lipid composition through post-burial degradation was assessed by incubation of potsherds dosed with cereal lipids at 35 °C in compost. The cooking experiments showed that only small quantities of cereal lipids are liberated, but additional lipid sources (meat) can increase the transfer of cereal biomarkers into the ceramic matrix. Anoxic degradation conditions allowed for twentyfold higher levels of alkylresorcinols and twofold higher levels of plant sterols after 20 weeks compared to oxic conditions. Therefore, samples from anoxic burial environments should be targeted and high sensitivity methods are a necessity to detect the trace amounts of cereal-specific biomarkers. To test the applicability of these biomarkers for archaeological pottery, organic residues from ten coarse ware vessels from an anoxic burial context at Vindolanda were analysed. Plant sterols and stanols were detected in three sherds, and two of the sherds also contained traces of alkylresorcinols.

The use of Lapita pottery: Results from the first analysis of lipid residues

Biomolecular and isotopic characterisation of absorbed organic residues have been performed on eight dentate-stamped and two plain Lapita potsherds from the site of Teouma, in Vanuatu. Lipid profiles associated with decorated pots are homogenous, suggesting that similar food types or mixtures of food types were placed in these vessels. This suggests a high degree of consistency in the use of Lapita decorated pots, irrespective of the morphological and stylistic variation of these vessels. Data obtained from single-compound isotope analysis are also not consistent with marine resources as potential food sources for Lapita vessels. The absence of such commonly consumed, ubiquitous and easily accessible resources in Lapita vessels suggests that these pots were not manufactured to be used for ordinary occasions and day-to-day food consumption. This is the first time tangible data related to the use of these vessels are provided to support this claim in addition to contextual inferences.

Strong bias towards carcass product processing at Neolithic settlements in northern Greece revealed through absorbed lipid residues of archaeological pottery

The emergence of agriculture in Greece denotes the start of the Neolithic in Europe, however, little is known about dietary practices in the region. Archaeobotanical and zooarchaeological remains indicate reliance on cereals and pulses, together with meat-based subsistence practices, including sheep/goat and pig husbandry. Preliminary investigations of dietary practices obtained through lipid residue analysis of pottery of a small number of sites in the region have confirmed primarily carcass products were processed. The weak evidence for dairy products contrasts with finding of dairy-based subsistence strategies in NW Anatolia, which is surprising given its close proximity. This paper aims to build on this earlier work to provide a more detailed model for the dietary changes throughout the region, both chronologically and spatially. To achieve this >900 potsherds from 11 sites spanning the Early (EN) to Late Neolithic (LN) periods from the north of Greece have been investigated using the lipid biomarker approach involving high temperature-gas chromatography (HT-GC), GC-mass spectrometry (GC-MS) and GC-combustion-isotope ratio MS (GC-C-IRMS) to determine the nature and origins of organic residues preserved in the fabric of pottery vessels. Lipid residue analysis of pottery vessels revealed ruminant and non-ruminant carcass fats comprise the majority of animal fat types identified, reflecting the high abundance of sheep/goat and pig in faunal assemblages. The emergence of dairying in northern Greece can now be dated to the site of EN/Middle Neolithic (MN) Ritini (5900/5700 - 5500 cal. B.C.E.), however, the frequency of dairy fat residues was low, overall, indicating that dairying was not intensively practised. The δ13C values of the fatty acids extracted from potsherds reflect a predominately C3 diet, however, in the EN and MN there is greater variation with some lipids exhibiting enriched δ13C values indicating a significant abundance of C4 plants in the ecosystem(s) covered by the study. Significantly, plant-derived n-alkanes (C22 to C34) detected in pottery vessels provide the first evidence for plant processing identified in lipid residues from ceramic vessels in Neolithic northern Greece, supporting the abundant archaeobotanical evidence for the processing of cereals and pulses.

Histoire de l’utilisation des laitages et de la persistance du gène de la lactase

This chapter covers archaeological and biological studies that have been insightful in the understanding of dairying history and the evolution of lactase persistence in humans. More specifically, we report results from lipid residue analyses on archaeological pottery vessels, ancient and modern human DNA and explain the biology and genetics of lactase persistence. Available data suggests that domesticated animals were used for milk consumption by humans early in the Neolithic, while contemporaneous populations seem to have been mainly lactose intolerant. This contrasts with modern European populations, which show a generally high proportion of lactase persistence.

GC–MS and HPLC-ESI-QToF characterization of organic lipid residues from ceramic vessels used by Basque whalers from 16th to 17th centuries

This work is focused on the chemical characterization of the organic residues preserved in ceramic fragments that are suspected to have been used by the Basque whalers in the period from 16th to 17th Century to store whale oil. The lipid profiling along with the identification of biomolecular markers, makes possible to obtain information on the contents of the vessels and on the transformations undergone by the organic substances during the burial process. To achieve this goal, two complementary analytical techniques, High-Performance Liquid Chromatography - Electrospray Ionization - Quadrupole Time-of-Flight Mass Spectrometry (HPLC-ESI-QToF) and Gas Chromatography-Mass Spectrometry (GC–MS), were used not only to study the archaeological materials but also fresh whale oil products used as reference materials with the aim to detect potential diagnostic biomarkers. HPLC-ESI-QToF was used to identify triacylglycerols (TAGs) and study their distribution in the various samples, whereas GC–MS after suitable derivatization provided the fatty acid (FA) profile together with the detection of degradation compounds (dicarboxylic and dihydroxy fatty acids) and biomarkers related to marine commodities (isoprenoid acids). Samples of blubber from 5 different whale species belonging to the genus Balaenoptera (fin, sei and minke whale), Megaptera (Humpback whale) and Phocoena (harbour porpoise) were used as reference materials together with a sample of whale oil of unknown origin. The chemical results confirm the archaeological data regarding the vessels use as containers to store whale oil and indicate the whales from Balaenoptera genus as the most probable source of the oil.

Wie bakterielle Moleküle unser Immunsystem manipulieren

Lipoproteins (Lpp) of Gram-positive bacteria play a crucial role in immune modulation. The degree of lipid acylation determines whether the TLR2/TLR1 or the TLR2/TLR6 signalling pathway is induced in the infected host. Both pathways differ significantly in the extent of innate immune activation. Commensal bacteria on our skin signal via the TLR2/TLR1 pathway thus inducing only a mild immune response while non-commensal bacteria signal via TLR2/TLR6 pathway thereby inducing a fulminant immune response. Consequently, the structure of the lipid residue in Lpp decides over immune tolerance or intolerance.

Preservation of uropygial gland lipids in a 48-million-year-old bird.

Although various kinds of organic molecules are known to occur in fossils and rocks, most soft tissue preservation in animals is attributed to melanin or porphyrins. Lipids are particularly stable over time-as diagenetically altered 'geolipids' or as major molecular constituents of kerogen or fossil 'geopolymers'-and may be expected to be preserved in certain vertebrate tissues. Here we analysed lipid residues from the uropygial gland of an early Eocene bird using pyrolysis gas chromatography mass spectroscopy. We found a pattern of aliphatic molecules in the fossil gland that was distinct from the host oil shale sediment matrix and from feathers of the same fossil. The fossil gland contained abundant n-alkenes, n-alkanes and alkylbenzenes with chain lengths greater than 20, as well as functionalized long-chain aldehydes, ketones, alkylnitriles and alkylthiophenes that were not detected in host sediment or fossil feathers. By comparison with modern bird uropygial gland wax esters, we show that these molecular fossils are likely derived from endogenous wax ester fatty alcohols and fatty acids that survived initial decay and underwent early diagenetic geopolymerization. These data demonstrate the high fidelity preservation of the uropygial gland waxes and showcase the resilience of lipids over geologic time and their potential role in the exceptional preservation of lipid-rich tissues of macrofossils.

MALDI‐FT‐ICR‐MS for archaeological lipid residue analysis

Organic residue analysis has become one of the major and rapidly developing fields in archaeological sciences. The analyses of lipid residues have produced several fruitful studies allowing a better understanding of past life-ways and economy, pottery function and agricultural practices. One potentially fruitful, yet under-utilized analytical method for the identification of archaeological lipid residues is MALDI-FT-ICR-MS. In this Perspective Special Feature, Ester Oras and colleagues demonstrate the suitability of MALDIFT- ICR-MS for archaeological organic residue analysis with four different archaeological samples and highlight its capability of identifying different intact, degraded and oxidized triacylglycerols from food residues. The article describes the analysis of samples of both food-crust and ceramic powder from vessels discovered from the 12th–13th century AD cemetery at Kukruse, Estonia. Dr. Oras is a Research fellow of, Analytical Chemistry, Institute of Chemistry, University of Tartu (Tartu, Estonia). Her research interests are organic residue analysis of prehistoric and early medieval material with her main focus on food residues, but she has also worked on adhesives, embalming resins, and isotope based migration studies.

MALDI-FT-ICR-MS for archaeological lipid residue analysis.

Soft-ionization methods are currently at the forefront of developing novel methods for analysing degraded archaeological organic residues. Here, we present little-used soft ionization method of matrix assisted laser desorption/ionization-Fourier transform-ion cyclotron resonance-mass spectrometry (MALDI-FT-ICR-MS) for the identification of archaeological lipid residues. It is a high-resolution and sensitive method with low limits of detection capable of identifying lipid compounds in small concentrations, thus providing a highly potential new technique for the analysis of degraded lipid components. A thorough methodology development for analysing cooked and degraded food remains from ceramic vessels was carried out, and the most efficient sample preparation protocol is described. The identified components, also controlled by independent parallel analysis by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS), demonstrate its capability of identifying very different food residues including dairy, adipose fats as well as lipids of aquatic origin. The results obtained from experimentally cooked and original archaeological samples prove the suitability of MALDI-FT-ICR-MS for analysing archaeological organic residues. Sample preparation protocol and identification of compounds provide future reference for analysing various aged and degraded lipid residues in different organic and mineral matrices.

QUANTUM CHEMICAL STUDY OF OXIDATION REACTIONS IN UNSATURATED HYDROCARBONS

In this article quantum-chemical calculations using DFT B3LYP/cc-pvdz method was used for the investigation of changes in thermodynamic functions of reactions of oxidation of unsaturated hydrocarbons such as heptane and heptadiene as low-molecular models of hydrocarbon residues of lipids. The effect of the position of the reaction center relative to the double bonds and conformations of double bonds on reactivity of the compounds in reactions of hydrogen abstraction by the hydroxyl radical, dioxygen accession and chain propagation were analyzed. By comparison of changes in thermodynamic functions of reactions it was shown that hydrocarbons with cis- conformations of double bonds are characterized with higher reactivity in reactions of hydrogen abstraction but peroxi-radicals of these conformers are more stable. The changes in thermodynamic functions of reaction of hydrogen abstraction for diene according to the calculation are smaller comparing with olefins. This is due to the difference in the stability of the radicals formed. The stability of hydrocarbon radicals of dienes in comparison with olefins is explained by their planar structure with electron density of unpaired electron delocalized between five carbon atoms. The emergence of such pentadienil-type radicals is the cause of a higher oxidation of dienes compared with olefins. The analysis of molecular structures of peroxi-radicals of dienes shows that after accepting dioxygen by hydrocarbon radical the isomerization takes place. According calculations it is preferable for the dioxygen molecule not to join with the central carbon atom from which the hydrogen atom has been abstracted but to attack the double bond joining with C2 carbon atom. During the isomerization the double bond moves to the center of the molecule forming thus the conjugated pair with the other double bond. Comparison of thermodynamic functions of reaction for cis- and trans- isomers shows that cis-trans isomerization is possible during the dioxygen accession to the hydrocarbon radical. These results are in good agreement with the experimental data published earlier.Forcitation:Novikova A.A., Soloviev M.E. Quantum chemical study of oxidation reactions in unsaturated hydrocarbons. Izv. Vyssh. Uchebn. Zaved. Khim. Khim. Tekhnol. 2017. V. 60. N 7. P. 14-20.

Lipid Residue Analysis of Archaeological Pottery: An Introductory Laboratory Experiment in Archaeological Chemistry

In this research-based experiment, students are introduced to the interdisciplinary field of archaeological chemistry by extracting and analyzing lipid residues absorbed in pottery. Reproduction archaeological pottery sherds are prepared by soaking ceramic fragments in individual or combinations of vegetable oils. Students crush and extract the absorbed residues, transesterify the recovered lipids to fatty acid methyl esters, and analyze the product by GC or GC/MS. Recovered residues are characterized by analysis of the major fatty acid peaks, as identified using reference standards or mass spectral databases. An archaeological context that links the sherds to the Minoan civilization is provided to students and must be considered to correctly identify the absorbed residue(s). The laboratory has been used in a topical archaeological instrumentation course that has attracted second to fourth year students majoring in chemistry, biochemistry, and anthropology, and museum studies minors. Pedagogically, the la...

Ligand chain length drives activation of lipid G protein-coupled receptors

Sphingosine-1-phosphate (S1P) is a lipid mediator that can activate five cell membrane G protein-coupled receptors (GPCRs) which carry a variety of essential functions and are promising drug targets. S1P is composed of a polar zwitterionic head-group and a hydrophobic alkyl chain. This implies an activation mechanism of its cognate receptor that must be significantly different from what is known for prototypical GPCRs (ie receptor to small hydrophilic ligands). Here we aim to identify the structural features responsible for S1P agonism by combining molecular dynamics simulations and functional assays using S1P analogs of different alkyl chain lengths. We propose that high affinity binding involves polar interactions between the lipid head-group and receptor side chains while activation is due to hydrophobic interactions between the lipid tail and residues in a distinct binding site. We observe that ligand efficacy is directly related to alkyl chain length but also varies with receptor subtypes in correlation with the size of this binding pocket. Integrating experimental and computational data, we propose an activation mechanism for the S1P receptors involving agonist-induced conformational events that are conserved throughout class A GPCRs.

Exploring the culinary uses of Santa María and Belén painted vessels from the Late Intermediate Period in Catamarca, Argentina

Santa María and Belén vessels from the Late Intermediate Period (10th–15th centuries AD) in Northwest Argentina are strongly associated to funerary uses as “urns”. However, multiple recent findings of these vessels in domestic non-funerary contexts point towards diverse utilities. In this paper we carried out an exploratory analysis and selected two case studies of Santa María vessels and one of a Belén vessel, all recovered in household floors from three sites in Catamarca, Argentina. In order to enquire into the potential culinary uses of vessels, lipid residues were extracted from the ceramic matrixes and gas chromatography–mass spectrometry (GC–MS) analyses were carried out to explore their origins. In all three vessels lipid residues were recovered and characterized. Two samples showed evidence of plant and animal lipid mixtures, while the third sample only had animal lipids. The culinary utility of these vessels was confirmed, and uses may have included storage or service of liquids or stews. These preliminary results provide insight into an alternative interpretation of Santa María and Belén vessel use.

New insights into the subsistence economy of the Eneolithic Dereivka culture of the Ukrainian North-Pontic region through lipid residues analysis of pottery vessels

The Dereivka site of the North-Pontic forest-steppe has been widely investigated because of its potential as a centre for horse domestication (Levine, 1990; Telegin, 1986). Despite the significant archaeological evidence available, Dereivka is considered a contradictory and complex site (Rassamakin, 1999: 143) due to a range of challenges connected with reconciling the various lines of available archaeological evidence. Consequently, a generally acceptable subsistence economic model has still to be developed, with contrasting theories remaining unresolved. This paper presents new results of organic residues analyses from the site. Forty potsherds were submitted to biomolecular and stable carbon and hydrogen isotope analyses and the results discussed in relation to previously published zooarchaeological evidence (Bibikova, 1986; Levine, 1999; Kaiser, 2010). The findings offer a further perspective on the overall subsistence economic strategies of the community, particularly in relation to the exploitation of the horse. Significantly, the biomolecular and stable carbon isotope results confirmed that Dereivka community consumed horse products predominantly, together with smaller proportions of ruminant and non-ruminant products. Interestingly, although ruminant adipose fats were recovered from some vessels, evidence of ruminant dairy product exploitation was insignificant, with only one residue displaying a possible ruminant dairy fat origin. Hydrogen isotope analysis of lipids was applied to investigate equine milk processing in pots (Outram et al., 2009) but these analyses did not offer significant new insights.

LATE MESOLITHIC NARVA STAGE IN ESTONIA: POTTERY, SETTLEMENT TYPES AND CHRONOLOGY

This paper gives a systematized overview of different Narva stage sites in Estonia, describing their artefactual and archaeozoological material, and environmental conditions. We demonstrate the diversity of Narva stage settlement types (sites on coastal river estuaries, coast, coastal lagoons, inland river banks and shores of inland lakes) and economy (marine, terrestrial/inland aquatic and mixed subsistence) in the region. A further site-based description of Narva pottery is also provided in order to exemplify the similarities and differences of this earliest pottery type in the eastern Baltic. We also present a comprehensive list of all currently available Narva stage radiocarbon dates from Estonia and Ingermanland (north-western Russia) according to which the Narva-type pottery in the northern part of its distribution area dates to the period c. 5200-3900 cal BC. Additionally, the issues of dating food crust, especially the high risk of reservoir effect offsets, are emphasized. We carried out a comparative study dating contemporaneous food crusts and plant remains and conducting lipid residue analysis employing combined methods of gas chromatography-mass spectrometry (GC-MS) and isotope ratio mass spectrometry (GC-C-IRMS, EA-IRMS). The results demonstrate the implications and importance of characterizing lipid residues so that samples with reservoir correction can at least be identified. Introduction The beginning of pottery use is a substantial and important innovation in the history of humankind. It must have brought along a new concept not only for storage and cooking, but also changes in technology and production skills, transport and trading. The earliest pottery known to date is from eastern Asia (China, Japan and Russian Far East and eastern Siberia) and goes back to the end of the Pleistocene (e.g. Nakamura et al. 2001; Kuzmin 2006; Boaretto et al. 2009; Shevkomud & Yanshina 2012, 53). In the Baltic Sea region pottery first appears, depending on traditions of archaeological periodization, in the Early Neolithic (Finland, Russia, Latvia, Lithuania, Belarus) or Late Mesolithic period (Sweden, Denmark, Germany, Poland, Estonia) (e.g. Loze & Lijva 1989; Girininkas 2005, 105 f.; Kriiska 2009, fig. 5; Piezonka 2012, 24). Earlier in Estonia the introduction of pottery was assigned to the Neolithic period, as with other Baltic States (e.g. Jaanits et al. 1982, 61; Lang & Kriiska 2001, 89). Since the adoption of pottery generated no significant change in the local settlement pattern, subsistence economy nor, presumably, in social organization, the new periodization of the Stone Age in Estonia proceeds from the process of the introduction of domesticates. The latter occurred more than a thousand years after the adoption of pottery and resulted in major shifts in the society, settlement and economy in both Estonia and neighbouring areas (Kriiska 2009, 167; Nordqvist et al. 2015, 143). Two major pottery traditions, divided into many sub-variants, can be distinguished in the Baltic Sea region. One of the traditions embraces pottery types that comprise large pointed- or rounded-bottomed pots and, in places, also small and shallow saucer-shaped vessels (see e.g. Hallgren 2004, fig. 1). The know-how of making such vessels spread from the late 6th millennium through the early 5th millennium cal BC (e.g. Loze 1988, 101; Hallgren 2004, 136 f.; Piezonka 2008, 76; 2012, 42; Girininkas 2009, 127; Kriiska 2009, 161; Jennbert 2011, 99; Pesonen et al. 2012). Five types of early pottery are traditionally distinguished in the Baltic Sea region: (1) Ertebolle in southern Scandinavia, northern Germany and northern Poland (e.g. Hallgren 2004, 135; Jennbert 2011); (2) Early comb ware (or Sperrings) in southern Finland and north-western Russia (e.g. Luho 1957; German 2002); (3) Saraisniemi 1 in northern Finland and in Karelia, the Kola Peninsula in Russia and northern Sweden (e.g. Torvinen 2000; Haggren et al. …