GC–MS ANALYSIS OF LIPIDS IN PREHISTORIC ROCK PAINTS AND ASSOCIATED OXALATE COATINGS FROM THE LOWER PECOS REGION,TEXAS*

The presumption that lipids were used as binders/vehicles in pictograph paints from southwestern Texas was tested using gas chromatography–mass spectrometry (GC–MS). A one‐step transesterification/derivatization procedure was used to convert bound and unbound fatty acids to fatty acid methyl esters for the analysis. Approximately 30 organic compounds were detected in the natural rock coating that encapsulates the paints, but there were no compounds unique to the paints. Moreover, the C_16:0 and C_18:0 concentrations in the paint samples were similar to the concentrations of these fatty acids in the rock coating, indicating that the compounds are native to the patina. Thus, lipid binders were either not used in the original paint mixture or have since degraded.     

Application of lipid biomarker analysis to evaluate the function of ‘slab-lined pits’ in Arctic Norway

Gas chromatography–mass spectrometry (GC–MS) and bulk carbon isotope determinations have been performed on samples (‘cemented organic residues’, charcoal, sediment and fire-cracked rock) excavated from 12 slab-lined pits from various locations in Arctic Norway to test the premise that these archaeological features were used for the extraction of oil from the blubber of marine mammals, such as seal, whale and walrus. A wide range of lipid compound classes were detected especially in the cemented organic residues and in the charcoal samples. The presence of long-chain unsaturated and isoprenoid fatty acids together with oxidation and thermal alteration products of unsaturated acids such as dicarboxylic acids, dihydroxyfatty acids and ω-(o-alkylphenyl)alkanoic acids suggests that these features were used for marine oil extraction at elevated temperatures. Notably the location of the hydroxyl groups in the dihydroxyfatty acids provides a record of the positional isomer of the precursor fatty acid and allows confirmation that 11-docosenoic (cetoleic) acid, the most abundant C_22:1 isomer in marine oil, was a major component of the original lipid. Further information was provided by the presence of long-chain fatty acyl moieties in surviving triacylglycerols and the presence of cholesterol. A fungal metabolite, mycose (trehalose), was found in all samples apart from a fire-cracked rock and points to microbiological activity in the pits. Bulk isotope analysis conducted on the ‘cemented organic residues’ is consistent with modern reference samples of blubber and oil from seal and whale. These data provide clear analytical evidence of the function of slab-lined pits in the archaeological record and suggest widespread exploitation of marine mammals for producing oil for heating, lighting and myriad other uses in the past.     

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.     

A NEW METHOD FOR EXTRACTION,ISOLATION AND TRANSESTERIFICATION OF FREE FATTY ACIDS FROM ARCHAEOLOGICAL POTTERY

This paper presents evidence for increased recovery of organic residues from archaeological pottery through use of a microwave‐assisted liquid chromatography protocol. C_16:0 and C_18:0 saturated fatty acids were obtained from archaeological potsherds recovered from nine Neolithic settlements in the Middle East dating between 4700 and 7300 cal bc , including materials that had not produced evidence for the survival of any lipid species through use of ‘conventional’ solvent extraction techniques. Compound‐specific isotopic analyses of C_16:0 and C_18:0 fatty acids in potsherd extracts subsequently revealed δ¹³C/δ¹²C compositions consistent with modern subcutaneous fats of wild boar and goats pastured on lands adjacent to the Jordan Valley, and residues from a modern pottery vessel used in the manufacturing of butter, cheese and yogurt in central Turkey. These results are presented as an illustration of capabilities of the microwave‐assisted recovery protocol. The reclamation of saturated and unsaturated fatty acids from archaeological pottery fragments recovered from a number of the earliest ceramic horizons in the Middle East is herewith reported, and the extraction methods and instrumental analytical techniques are described.     

EXPERIMENTAL EVIDENCE FOR THE PROCESSING OF MARINE ANIMAL PRODUCTS AND OTHER COMMODITIES CONTAINING POLYUNSATURATED FATTY ACIDS IN POTTERY VESSELS*

ω‐(o‐Alkylphenyl)alkanoic acids are known to form in vitro from triunsaturated fatty acids following protracted heating. These compounds have recently been identified in absorbed lipid extracts obtained from archaeological pottery vessels, providing a potentially valuable new class of indicator for the processing of commodities, such as marine oils, which contain high abundances of polyunsaturated fatty acids. Experiments were conducted to assess whether ω‐(o‐alkylphenyl)alkanoic acids form when pure compounds and complex triacylglycerol mixtures are heated with a fired clay. The results demonstrate that ω‐(o‐alkylphenyl)alkanoic acids are only produced following heating of unsaturated fatty acids (tri‐, di‐ and monounsaturated species) or complex unsaturated fatty acyl lipids at around 270°C. Heating saturated fatty acyl lipids does not yield ω‐(o‐alkylphenyl)alkanoic acids. Thus, when searching for evidence for the processing of marine derived animal fats in archaeological pottery vessels, it is essential that: ω‐(o‐alkylphenyl)alkanoic acids of carbon length C₁₈ and C₂₀ should be present, ideally with C₂₂ also detectable (if only in trace abundances), together with at least one of the three isoprenoid fatty acids (phytanic, pristanic or 4,8,12‐tetramethyltridecanoic acid).     

GC–MS ANALYSIS AND FATTY ACID RATIOS OF ARCHAEOLOGICAL POTSHERDS FROM THE WESTERN GREAT BASIN OF NORTH AMERICA*

The study of organic residues in archaeological pottery has focused on fatty acids due to their relative stability and longevity. However, even these compounds are subject to degradation, which makes assignment of residues to original foods problematic. This paper suggests that the use of ratios of fatty acids that degrade at roughly the same rate can be useful to identify very general categories of foods. It compares independent information on pot function based on ethnography and engineering/technological studies to that reconstructed based on extracted fatty acid ratios. The results support the notion that Great Basin pots were used primarily to boil seeds and that pot shape and pot function were related.     

GC/MS ANALYSIS OF FATTY ACIDS FROM ANCIENT HEARTH RESIDUES AT THE SWAN POINT ARCHAEOLOGICAL SITE*

Gas chromatography – mass spectrometry (GC/MS) is used to analyse soil residues from hearth sites in Swan Point, Alaska. An experimental hearth is also prepared by burning animal bones to study the characteristics of bone fires and the lipid residues that they leave embedded in soil. Hearth soil samples are derivatized with HCl in methanol to convert bound and unbound fatty acids into methyl esters. Concentrations of saturated fatty acid methyl esters in the samples are determined and their ratios analysed. The fatty acid patterns found in ancient campfires are consistent with burning bones of large ruminants as well as monogastric herbivores.     

ANALYSIS OF THE FATTY DEBRIS FROM THE WRECK OF A BASQUE WHALING SHIP AT RED BAY,LABRADOR

Analysis of fatty material from barrels in a submerged wreck of a whaler in Labrador water showed it to contain free fatty acids, including some higher fatty acid, consistent with whale oil and certainly of marine origin. Conversion to adipocere was extensive but not complete.     

Human soft tissue preservation in the Cova des Pas site (Minorca Bronze Age)

The preservation process of soft tissues in an archeological context is mainly unknown because they occur only in truly exceptional situations. The Cova des Pas is a Bronze Age site in Minorca where the special conditions enabled the preservation of some soft tissues associated with 66 individuals. This finding allows the study of the preservation process that took place by means of the analysis of the histological and chemical characteristics of the tissues. Our results show that the preservation mechanism was the adipocere, because the fatty acids profile shows higher concentration of saturated than unsaturated fatty acids. The evidence indicates that the kind of funerary ritual and the environmental conditions favored this preservation.     

Bennett’s wallaby (Macropus rufogriseus) marrow quality vs quantity: evaluating human decision-making and seasonal occupation in late Pleistocene Tasmania

A pilot study on the fatty acid composition of the bone marrow, brain tissue and muscle (flesh) of the medium-sized marsupial the Bennett’s wallaby (Macropus rufogriseus), aimed to explore the mechanisms behind the dominance of this human prey species in late Pleistocene southwest Tasmania. In particular it was designed to investigate why humans frequently selected and transported the wallaby hindquarters, breaking open their longbones and metatarsals to expose the marrow cavity, when these animals could easily have been transported whole. Initial results indicated that the percentage of unsaturated fatty acids particularly palmitoleic acid (C_16:1) and oleic acid (C_18:1) in the bone marrow increased distally away from the body core temperature or the heart, towards the hands and feet. There is a corresponding decrease in the percentage of the saturated fatty acids, particularly palmitic acid (C_16:0) and stearic acid (C_18:0). This is similar to the pattern identified in placental artiodactyls (even-toed ungulates) and rodents. The preliminary study was then expanded to a year-long seasonal experiment involving wallabies from different Tasmanian environmental and climatic zones to see if this can better help explain the apparent seasonal occupation of caves indicated from recent Bennett’s wallaby teeth annuli (Skeletochronological studies). It was found that the Bennett’s wallaby remained a relatively stable and dependable resource throughout the year, with environmental, altitudinal and seasonal variability having only a minor effect on the fatty acid composition or nutritional quality of their bone marrow, muscle and brain tissue. Interestingly, the animals had increased desirable unsaturated fatty acids in their marrow, muscle and brain during the winter and spring with decreased amounts during the summer and autumn. This lends support to an earlier Kidney Fat Index (KFI) analysis for the Bennett’s wallaby which recorded significant seasonal differences within and between regions, indicating an environmental effect on overall body condition. Fatty acid analysis of the Common wombat Vombatus ursinus, the Tasmanian pademelon Thylogale billardierii, and the Forester or Eastern Grey kangaroo Macropus giganteus were also included to ascertain why these animals remained constant, although minor, prey species throughout the late Pleistocene. While the results from these animals were similar to the Bennett’s wallaby, there were some unexpected and surprising patterns that require further investigation. All of these fatty acid analyses have important implications for understanding human decision-making and occupation of southwest Tasmania during the Last Glacial Maximum, for modern dietary and palaeodietary benefits, and for understanding the physiology of these unique Australian marsupials.     

A COMPARISON OF METHODS FOR ESTABLISHING FATTY ACID CONCENTRATION GRADIENTS ACROSS POTSHERDS: A CASE STUDY USING LATE BRONZE AGE CANAANITE AMPHORAE*

Canaanite amphorae fragments in fabrics believed to be associated with the bulk transport of vegetable oils from the fourteenth‐century BC site of Amarna, Egypt, have been examined. A comparison is made between solvent extraction, saponification and the use of TMTFTH (m‐trifluoromethylphenyl)trimethylammomum hydroxide, used here for the combined extraction and derivatization of ceramic‐absorbed organic residues. Extracts were studied using gas chromatography and gas chromatography‐mass spectrometry. The extraction of fatty acids from small ceramic samples has established concentration gradients of absorbed organic matter from the inner to the outer surfaces of the sherds.     

印谱古籍中的印油老化研究

存世印谱中比较常见的一类病害是印蜕中印油老化导致的渗化问题。研究表明,印油渗化不仅会在印蜕四周形成黄色或棕色的油渍,而且会导致纸张pH值降低,影响印谱的长久保存。通过对印泥中常用的蓖麻油、菜籽油、茶油、调和印油等六种油脂进行模拟老化实验,发现油脂老化后普遍存在变色、pH值下降和渗化问题。采用气相色谱-质谱联用技术分析老化前后的油脂样品,发现油脂在老化过程中,不饱和脂肪酸含量降低,饱和脂肪酸占比增加,并形成了壬二酸等脂肪族羧酸。油脂老化过程中的成分变化,导致了变色、酸化、渗化等一系列问题。在印谱实物样品中也发现了饱和脂肪酸。在印谱的保护工作中,应对油脂的老化进行有针对性的处理。     

固相微萃取-气相色谱-质谱在《江友渚等七挖书画轴》挥发性有机化合物分析中的应用

文化遗产的气味不仅反映了文物的历史信息,还对文化遗产自身产生持续性的影响,因此了解文化遗产的气味对其价值研究和当代保护都具有重要意义。本工作采用固相微萃取-气相色谱-质谱技术对中国国家博物馆藏《江友渚等七挖书画轴》天头用纸散发的气体进行了采集、分离和分析。结果表明：与纸张降解相关的标志物(如脂肪酸、醛、酮等)得到有效采集;而其他生物标记物(如沉香螺醇)的检出也反映了该书画轴的历史保存环境信息。此方法对该件书画轴的历史价值认知和科学保护具有重要作用。     

CHEMICAL COMPOSITION OF A BOG BODY ADIPOCERE

Chemical analyses were carried out on adipocere obtained from a bog body recovered from a peat bog at Meenybradden, County Donegal, Ireland. Chromatographic (thin-layer chromatography and gas chromatography) and mass spectrometric analyses, combined with microanalytical chemical transformations, have yielded detailed compositional information. An absence of intact triacylglycerols, diacylglycerols and monoacylglycerols indicates that hydrolysis is complete. Consequently the adipocere is composed mainly of fatty carboxylic acids. The high proportion of palmitic and stearic acids, together with depleted oleic acid content, indicates that extensive reduction and, possibly, β-oxidation have occurred during burial in the peat bog. Hydration of the double-bond in oleic acid has also occurred, as is shown by the formation of 10-hydroxystearic acid. The monoenoic fatty carboxylic acids that are present, composed mainly of C₁₈ and C₁₆ compounds, comprise a mixture of positional isomers. The results are compared and contrasted with those obtained from previous studies of the lipid composition of other adipoceres and cadavers of archaeological interest.     

Subsistence practices and pottery use in Neolithic Jordan: molecular and isotopic evidence

This paper presents direct evidence of subsistence practices and pottery use at a Late Neolithic site at al-Basatîn, northern Jordan. Measurable concentrations of C_16:0 and C_18:0 were recovered from 8 of 10 archaeological pottery fragments through use of a microwave-assisted silica gel and aminopropyl solvent protocol developed for the isolation and concentration of free fatty acids in marine sediments. Subsequent isotopic analysis of the surviving C_16:0 and C_18:0 saturated fatty acids revealed ∂¹³C ratios consistent with those of adipose fats of ruminant and non-ruminant animals pastured on lands adjacent to the Jordan Valley. The high recovery of diagnostic compounds from the al-Basatîn material is discussed in context of a wider examination of the initial development and use of pottery in the Fertile Crescent, and the emerging debate concerning the efficacy of stable carbon isotope values in characterizing organic residues embedded in pottery fragments recovered from the earliest ceramic horizons in the Middle East and Europe.     

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.     

Microstructural,chemical and isotopic evidence for the origin of late neolithic leather recovered from an ice field in the Swiss Alps

Archaeological leather samples recovered from the ice field at the Schnidejoch Pass (altitude 2756 m amsl) in the western Swiss Alps were studied using optical, chemical molecular and isotopic (δ¹³C and δ¹⁵N of the bulk leather, and compound-specific δ¹³C analyses of the organic-solvent extracted fatty acids) methods to obtain insight into the origin of the leather and ancient tanning procedures. For comparison, leathers from modern native animals in alpine environment (red deer, goat, sheep, chamois, and calf/cow) were analyzed using the same approach. Optical and electron microscopically comparisons of Schnidejoch and modern leathers showed that the gross structure (pattern of collagen fibrils and intra-fibrils material) of archaeological leather had survived essentially intact for five millennia. The SEM studies of the hairs from the most important archaeological find, a Neolithic leather legging, show a wave structure of the hair cuticle, which is a diagnostic feature for goatskins. The variations of the bulk δ¹³C and δ¹⁵N values, and δ¹³C values of the main fatty acids are within the range expected for pre-industrial temperate C₃ environment. The archaeological leather samples contain a mixture of indigenous (from the animal) and exogenous plant/animal lipids. An important amount of waxy n-alkanes, n-alkan-1-ols and phytosterols (β-sitosterol, sitostanol) in all samples, and abundant biomarker of conifers (nonacosan-10-ol) in the legging leathers clearly indicate that the Neolithic people were active in a subalpine coniferous forest, and that they used an aqueous extract of diverse plant material for tanning leather.     

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.     

THE CHARACTERIZATION OF PAINTS AND WATERPROOFING MATERIALS FROM THE SHIPWRECKS FOUND AT THE ARCHAEOLOGICAL SITE OF THE ETRUSCAN AND ROMAN HARBOUR OF PISA (ITALY)*

This work presents the results of a diagnostic survey on the shipwrecks from the archaeological site of the ancient harbour of San Rossore (Pisa, Italy). The original waterproofing, caulking and painting materials were characterized by Fourier transform infrared spectroscopy (FTIR), gas chromatography – mass spectrometry (GC–MS), energy‐dispersive X‐ray microanalysis (EDX) and X‐ray diffraction (XRD). The major constituents of the waterproofing and caulking materials detected on the planks were tricyclic abietanes showing a high degree of aromatization. These compounds are indicative of a pitch obtained from the wood of trees of the Pinaceae family. The analysis of the organic components of the paint samples revealed diterpenoid acids characteristic of Pinaceae resins together with linear long‐chain alcohols and fatty acids, highlighting the presence of beeswax. Noticeably, the characteristic odd carbon number alkanes that are normally present in beeswax were not detected. The EDX and XRD analyses showed that hematite, calcite, cerussite and kaolinite were employed as pigments to paint the ships’ hulls.     

Identification of maize in absorbed organic residues: a cautionary tale

Starchy grains are an essential part of human diet in most agricultural groups, and are attributed an important role in the development of complex societies. Maize is a starchy grain domesticated in Mesoamerica that was an important foodstuff throughout the Americas before Contact, and around the world afterwards. An experimental study of the degradation of maize lipids suggests that the unsaturated fatty acids comprising the majority of maize lipids degrade rapidly, producing a virtually unidentifiable organic residue after 3 months deposition. Compounds in maize lipids decompose variably, depending upon depositional environment, making calibration of organic residue degradation impracticable. Variable decomposition of components of absorbed organic residues makes a wider range of improved experimental studies important, and suggests that identifying maize, and by implication starchy grains, from chemical analysis of absorbed organic residues requires a wider range of approaches than those previously attempted.