Journal Pre-proofs

Extending the inter-Adriatic trade of bitumen beyond the fifth millennium

BCE

Antonio Pennetta, Daniela Fico, Giacomo Eramo, Italo Maria Muntoni,

Giuseppe Egidio De Benedetto

PII: S0146-6380(20)30048-6
DOI: https://doi.org/10.1016/j.orggeochem.2020.104013
Reference: OG 104013

To appear in: Organic Geochemistry

Received Date: 31 October 2019

Revised Date: 25 February 2020
Accepted Date: 6 March 2020

Please cite this article as: Pennetta, A., Fico, D., Eramo, G., Maria Muntoni, I., Egidio De Benedetto, G.,
Extending the inter-Adriatic trade of bitumen beyond the fifth millennium BCE, Organic Geochemistry (2020),
doi: https://doi.org/10.1016/j.orggeochem.2020.104013

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Extending the inter-Adriatic trade of bitumen beyond the fifth millennium BCE

Antonio Pennettaa*, Daniela Ficoa, Giacomo Eramob, Italo Maria Muntonic, Giuseppe Egidio De

Benedettoa

a Laboratorio di Spettrometria di Massa Analitica ed Isotopica, Dipartimento di Beni Culturali,

Università del Salento, Lecce, Italy

b Dipartimento di Scienze della Terra e Geoambientali, Università degli Studi di Bari "Aldo Moro",

Bari, Italy
c Soprintendenza Archeologia Belle Arti e Paesaggio per le province di Barletta-Andria-Trani e

Foggia, Foggia, Italy

*Corresponding author: E-mail: antonio.pennetta@unisalento.it

ABSTRACT

Grotta dei Cervi (Apulia, Southern Italy) is the most important European figurative art site for its

Neolithic red and brown paintings found in the various galleries of the cave. It was frequented with

particular intensity between an advanced phase of the Early Neolithic (first half of the sixth

millennium BCE) and continued up to the end of the Bronze Age (last centuries of the second

millennium BCE). Among the more than thirty thousand finds, some ceramic shards had black

residues on them. These residues were sampled so that molecular and isotopic investigations could

be carried out. GC–MS-MS and EA–IRMS after fractionation of the samples allowed us to identify

bitumen on some archaeological potsherd remains and to identify its geographic source by

comparison with geological samples from Majella and Selenicë, two deposits located respectively

in Italy and Albania and known to have been exploited since Neolithic times. All the bitumens from

Porto Badisco correlate each other and with the Selenicë geological samples. Isotopic fingerprints

strongly support the biomolecular evidence and confirm the Albanian origin of the archaeological

bitumen. The data presented herein provide the first evidence of export and trade of raw bitumen

across the Adriatic Sea from Selenicë to Apulia during the Neolithic age thus bringing back to the

fifth millennium BCE the transport of bitumen across the Adriatic Sea in the central Mediterranean

area.

Keywords: Bitumen; Biomarkers; GC–MS-MS; EA–IRMS; Neolithic age; Albania.

1. Introduction

Bitumen, like amber (van der Werf et al., 2016) or flint, is considered a primary proxy of

trading patterns during prehistory. In fact, it occurs widely but the relevant mines or seepages are

well localized. Rullkötter and Nissenbaum (1988) and Connan and Dessort (1989) firstly analysed

archeological bitumens and succeeded in determining its source in balms of Egyptian mummies
using molecular criteria. The sterane and hopane biomarkers in the balms, correlated with Dead Sea

asphalt, proving sustained trade between Palestine and Egypt cultures. Since then, different

archeological bitumens have been provenanced as reviewed by Connan (1999) and Schwartz and

Hollander (2008). Less studied, possibly due to their less common occurrence, are archaeological

bitumens excavated in the central Mediterranean area although their possible occurrence in Bronze

age excavations has been reported (Guglielmino, 2012). Archeological findings from Monopoli and

Torre Santa Sabina excavation were provenanced using isotopic and biomarker analyses and their

Albanian origin was demonstrated (Faraco et al., 2016).

The Grotta dei Cervi is situated in Porto Badisco, Italy (Supplementary Fig. S1). It is part of

a complex system extending over the northern part of the Cervi Bay, known since the sixteenth

century as Porticello, in the Otranto channel along the coast of the Salento peninsula between Capo

d’Otranto and Santa Cesarea Terme. The cave complex includes a number of galleries branching

out in different directions within the limestone formations that characterise the area, along tectonic

fracture joints. The overall development of the cave is almost 2.5 km, at an average depth of ca. 25

m from the surface. Since its discovery in 1970 by the “Pasquale de Lorentiis” Salento

Speleological Group from Maglie (Lecce, Italy), various research campaigns have been carried out

in the Grotta dei Cervi. The cave is the most important European figurative art site for its Neolithic

red and brown paintings found in the various galleries of the cave (Graziosi, 1980). The paintings of

the Grotta dei Cervi are concentrated in three, almost parallel, corridors running approximately

north–south (with a particular concentration in corridor 2). In the entrance areas, and in the other

galleries without paintings or graffiti, a remarkable number of archaeological finds, structures such

as walls and circles built using the dry-wall technique and human depositions, were discovered both

on the surface and in the deposits investigated during various excavation campaigns carried out

from 1970 to 1997.

The prehistoric complex of the site was frequented with particular intensity between an

advanced phase of the Early Neolithic (first half of the sixth millennium BCE) and continued up to
the end of the Bronze Age (last centuries of the second millennium BCE). However, the various

excavations carried out have not made it possible to adequately set the paintings in relation to the

Holocene archaeological levels with evident cult, ritual and funerary, characteristics. Beneath the

Holocene levels represented by brown soils, there are levels of red earth of aeolian deposition,

containing the remains of still unpublished industries dating to the Upper Palaeolithic, the period to

which the first human visitations of the karst system are dated.

The recent systematic re-examination, carried out in collaboration with the University of

Salento, of the over 34,000 finds from the first 1970–1971 excavations by Felice Gino Lo Porto, at

that time the Superintendent of the Archaeological Heritage Office of Puglia, has confirmed the

long history of human use of the karstic complex (Aprile et al., 2017). In this paper, new

archaeological samples obtained from the Neolithic site of Grotta dei Cervi at Porto Badisco (in the

southeast of Italy) have been studied and the relevant result permitted to extend the inter Adriatic

trade of bitumen up to fifth millennium BCE.

2. Material and methods

2.1. Sampling

Five archaeological potsherds coming from the 1970–1971 excavation in Grotta dei Cervi of

Porto Badisco were selected for bitumen analysis (Fig. 1 and Supplementary Fig. S2) on the basis

of the macroscopic aspects of the samples. The samples were coded as Porto Badisco (PB). They all

show a thin layer of darkish material, probably organic matter (OM), along the fractures (PB1, PB2,

PB4, PB5), probably used to repair broken pieces, or on the outer surfaces (PB3). PB1 is one of

three fine Middle Neolithic body sherds coming from one of the areas accessible from Entrance D

and E. PB2 and PB5 are two typical handles of pottery in the Serra d’Alto style, the former (PB2)

discovered in one of the painted corridor in July 1970 and the latter (PB5) from Cavity E (from –

4.20 m to –4.40 m below the vault). PB3 is the bottom of a Neolithic vessel from Cavity A (from –
4.60 m to –4.80 m below the vault, recovered on April 7, 1970). PB4 is a body sherd, probably

Neolithic, from Cavity E (from –2 m to –2.20 m below the vault, recovered on April 23, 1970). The

layers were sampled along the border, by gently scraping them off with a scalpel, trying to avoid

including any ceramic material, as in this case it was hypothesized that the bitumen was used as a

glue. Generally, 10–100 mg of powder was recovered for geochemical analysis.

2.2. Sample preparation

The samples were studied using the standard procedures outlined in Faraco et al. (2016). In

summary, the samples were extracted with dichloromethane (DCM) and the asphaltenes were

precipitated from the extract using pentane and separated via centrifugation for 5 min (2000 rpm).

The precipitate was washed at least 3× with fresh pentane and weighed. The deasphalted pentane

solution, after concentration, was loaded onto an activated silica column (Sigma-Aldrich) to

separate saturated hydrocarbons (pentane), aromatic hydrocarbons (pentane/DCM, 70:30, v/v) and

the polar fraction (DCM/methanol). After fractionation and solvent removal, the saturated

hydrocarbon fraction was dissolved in hexane for analysis.

2.3. GC–MS and GC–MS-MS analysis

GC–MS analyses were performed using an Agilent 6890N gas chromatograph interfaced

with an Agilent 5973inert mass spectrometer. The GC was fitted with a DB-1 fused silica column

(60 m). High purity He was used as carrier gas with a constant flow rate of 1.5 mL/min. Samples

were injected at 280 °C in splitless mode (split vent opened after 0.5 min) using a 7683B

autosampler. The GC oven was programmed from 40 (2 min) to 200 °C at 20 °C/min, then to 310

°C at 2 °C/min with a final hold time of 35 min. Saturate fractions were analysed in full scan (50–

550 Da) and in single ion monitoring (SIM) mode (m/z 177, 183, 191, 205, 217, 218, 221, 231,

259).
 A Bruker SCION-456GC gas chromatograph equipped with PAL autosampler and coupled

to a Bruker SCION TQ triple quadrupole mass spectrometer was used with a DB–1 fused silica

column (60 m × 0.25 mm i.d., 0.25 µm thickness). The injector temperature was 260 °C. The oven

temperature program was: 50 °C (2 min) to 150 °C at 20 °C/min, then to 312 °C (held 15 min) at

1.5 °C/min. Electron impact (EI) ionization mode (70 eV) was used and He was the carrier gas at a

constant flow (1.2 mL/min). Ar was used as the collision gas at 2.0 mbar and 10 V collision energy.

1.0 µL of sample was injected in splitless mode. Although, absolute concentrations are preferred to

calculate peak ratios, but considering that the same MS-MS transitions were involved, peak areas

were used for calculation.

2.4. Stable carbon isotopic analysis

To determine the stable carbon isotope composition of asphaltenes, an aliquot (ca. 0.1 mg)

was analyzed on an Elementar IsoPrime 100 isotope ratio-mass spectrometry (IRMS) instrument

(IsoPrime Ltd., Cheadle Hulme, UK) coupled to an N-C-S elemental analyzer (Elementar Vario

Pyro Cube EA CNS; Elementar Analysensysteme GmbH, Hanau, Germany). The elemental

analyzer combustion tube was at 1020 °C and the reduction tube at 850 °C. He was at 230 mL/min

and O2 at 35 mL/min. He in the dilutor was at 4 bars and the CO2 reference gas at 4 bars. The

reference materials for determination of the stable carbon isotopic values (δ13C) were IAEA-CH-7

polyethylene (–32.151‰) and NBS 18 calcite (–5.014‰). The δ13C values were calculated relative

to Peedee Belemnite (PDB) as the mean of three measurements, the standard deviation not

exceeding 0.1‰.

3. Results and discussion

3.1. Bulk properties

 The archaeological samples from Porto Badisco were scraped from five different finds

(Table 1). As the amount of the available samples was from 6 to 100 mg, only three of them (PB2,

PB3 and PB4) gave reliable data for MS-MS determination of hopanes and steranes. The other two

samples (PB1 and PB5) gave dubious data, with very low signal/noise ratio for both steranes and

hopanes, so they are only described briefly. The extractable organic matter (EOM) in the PB1 and

PB5 samples was about 5 wt% so the bitumen content is very limited. Moreover, it was not possible

to sample the potsherds more extensively. The other samples had an EOM of 25–31 wt% , hence

PB2, PB3, and PB4 are mixtures of bitumen with mineral matter. Most bituminous mixtures contain

mineral matter, as commonly observed in most archaeological samples (Connan et al., 2013): a

portion of the mineral matter comes from the potsherd itself because of in-depth scraping of the

bituminous mixtures affecting the potsherd.

Comparison of bulk composition properties (Table 1) shows that the pentane-soluble

fraction is dominated by the saturated fraction, whereas the aromatic and the polar fractions are

somewhat limited. The accuracy of these values is low, due to the small amount of EOM: as a

result, they cannot be considered as significative for comparison with geological samples.

3.2. Biomarkers: gas chromatography–mass spectrometry (GC–MS and GC–MS-MS)

The assessment of bitumen origin has been carried out by using molecular tools, especially

sterane and terpane distribution patterns, which are useful for inferring genetic patterns and

highlighting bitumen sources (Peters and Moldowan, 1993; Connan et al., 2004; Peters et al., 2005).

The data were gained mainly by GC–MS-MS employing the transitions M+∙ → 217 for steranes and

M+∙ → 191 for hopanes (Faraco et al., 2016). Based on the distribution of C15+ saturated

hydrocarbons, all archaeological samples are biodegraded, roughly to level 6 on the Peters and

Moldowan scale (Peters and Moldowan, 1991). Fig. 2 shows a typical gas chromatogram of

saturated hydrocarbons from the archaeological bitumens (sample PB2): the chromatogram is

characterized by an absence of n-alkanes, but terpanes are well preserved and m/z 191 mass
chromatograms still give a good impression of the hopane distribution together with that of the

minor tri- and tetracyclic terpanes (Supplementary Fig. S3).

Tricyclic and tetracyclic terpanes are widespread in oils and bitumens. The formers are

found in both lacustrine and marine source rocks whereas the latter suggest a carbonate or evaporite

depositional environment. Hence, bitumens from carbonate source rocks can be distinguished by

high C24 tetracyclic terpane/C23 tricyclic terpane (Tetra24/T23) and low C24 tricyclic terpane/C23

tricyclic terpane (T24/T23) ratios. All our archaeological samples share these characteristics with

both Italian and Albanian bitumens (Table 2) so these ratios are not helpful for differentiating

bitumen sources.

3.2.1. Hopanes

Hopanes originating from bacterial (prokaryotic) membrane are present in all archaeological

samples and have a similar fingerprint (Supplementary Fig. S4). Whereas tri- and tetracyclic

terpanes were quantified using GC–MS, hopanes were quantified using GC–MS-MS data in order

to avoid errors from coelution: Fig. 3 shows the C27 and C29–35 mass chromatograms of samples

PB2, PB3, PB4 where all the peaks used for bitumen correlations are labelled. Variations in some

parameters are clearly dominated by maturity, others are dominated by source input and many are

affected by both source and maturity. The hopane/moretane ratio is an indicator of thermal maturity

whereas the 7α(H),21β(H)-30-norhopane/17α(H),21β(H)-hopane (H29/H30) ratio or the

homohopane distributions are more useful for source correlation. To the latter family belongs the

Ts/Tm ratio. Archaeological samples have maturity parameters such as 17α(H),21β(H)-30-

norhopane/17β(H),21α(H)-30-norhopane (H29/M29) and C30 17α(H),21β(H)–

hopane/17β(H),21α(H)-hopane (H30/M30) similar to those recorded for two of the Selenicë and

one of the Majella samples. These samples also have a similar H29/H30 ratio which is greater than

1.0 suggesting an anoxic carbonate or marl source rocks for the bitumens. Also, the 18α-22,29,30-

trisnorneohopane/22,29,30-trisnorhopane ratio (Ts/Tm) is low for all our samples and

characteristics of bitumens from carbonate source rocks, however archaeological Grotta dei Cervi

and geological Albanian bitumen samples also similar values for this parameter (Table 2).

Another common feature of Italian and Albanian bitumens is the decrease in abundance with

increasing carbon number of hopane homologues from C31 to C35. However, there are important

differences when considering the relative content of C33 and C34 isohopanes and the presence of C33

and C34 30-norhopanes in the bitumens: these molecules are more resistant to biodegradation and

the relevant ratios with respect to regular homohopanes (Table 3) are high for the more biodegraded

SE3 samples. Hexahydrobenzohopanes are considered diagnostic of bitumen from sulfur-rich

carbonate-anhydrite source rocks. They have been determined by GC–MS-MS and their ratios to

regular homohopanes show a great similarity among archaeological and some of the Albanian

samples (SE1 and SE2).

3.2.2. Gammacerane

Fig. 3 shows the hopane profile of the archaeological sample and one of the Selenicë

geological samples suggesting an Albanian origin for the archaeological bitumens. A high

gammacerane content is consistent with a carbonate source rock. Previous workers (Mattavelli and

Novelli, 1990) have suggested that the majority of the Italian oils and bitumens have a carbonate

source-rock affinity. On considering the biomarker ratio G/H29, which appears less affected by

biodegradation, it is clear that the Majella bitumens have significantly higher gammacerane

contents than the values recorded for the archaeological samples. In contrast, SE1 and SE2 have

quite similar G/H29 values (Table 2). Fig. 4 shows the plot of this biomarker ratio vs Tetra34/T23:

the three Porto Badisco samples are quite similar to SE1 and SE2, and are thus consistent with the

inferences drawn from the other hopane profiles. The samples from Majella – other geological

bitumens exploited in the Neolithic period – are rather different.

3.2.3. Steranes
 Fig. 5 shows the chromatograms relevant to the transitions m/z 372 → 217, m/z 386 → 217,

m/z 400 → 217, and m/z 414 → 217 for C27, C28 and C29, respectively, where both diasteranes and

regular steranes can be recognized. Regular sterane distributions of all archaeological samples show

a marked predominance of C29 steranes and lower proportions of C28 and C27 steranes (Table 2). It

is not possible to ascertain the organic matter inputs because of the steranes are biodegraded,

nonetheless the relevant profiles can be used to help identify the provenance of the archaeological

bitumens: in the Porto Badisco samples, the C29 steranes (60–64%) are more prominent followed by

C27 steranes (22–28%) and C28 steranes (12–15%) as observed for the Selenicë samples SE1 and

SE2.

3.3. Carbon isotopic composition of C15+ saturated hydrocarbons

Isotopic data (δ13C and δ34S) on asphaltenes are considered as source indicators (Connan and

Nishiaki, 2003) and therefore characteristic of the bitumen origin. The isotopic data (Table 3) show

that PB2, PB3, and PB4 have δ13Casph equal to –29.3‰, –29.3‰ and –29.1‰ respectively, so

clearly have the same isotopic signature. As for sulfur, the recorded values varied from –23.2‰ to –

24.2‰. Fig. 6 plots these data along with those recorded for the Majella and Selenicë samples

supporting an Albanian provenance for the Porto Badisco bitumens.

Comparison of these figures and data in Table 3 allows, in fact, the conclusion that all the

bitumens from Porto Badisco correlate with each other and with the Selenicë geological samples.

The outstanding differences are the significant amount of gammacerane and the almost absence of

diasteranes in the Majella samples. On considering both isotopic and biomolecular evidence, it is

possible to conclude that only SE1 and SE2 (coming from the same pit) are compatible with the

bitumens extracted from the archaeological findings (Supplementary Fig. S5).

4. Discussion
 The study of the ceramic material from Grotta dei Cervi cave (De Pascale and Muntoni,

2018; Ingravallo et al., 2019) shows the existence of a network of long-distance interactions

involving Neolithic communities, mainly from the Middle Neolithic phases, of the southeast Italy.

Early and Middle Neolithic sea voyaging, involving crossings of 100 km or more, are well

documented in the Mediterranean area (Freund and Batist, 2014; Zilhão, 2014). Contacts between

local Neolithic communities and Dalmatian coasts have been just previously suggested by other

scholars (Spataro, 2009; Colombo and Tozzi, 2013). Moreover, the use of the bitumen from the

Majella is well documented in Early and Middle Neolithic sites in Abruzzo, such as Catignano,

Sant’Angelo and Piccioni caves (Nardella et al., 2019).

The provenance of bitumen used to repair fractures from Selenicë specifically confirm the

long-distance interaction back to the fifth millennium BCE. An extended network of exchange

during the Neolithic period, mainly in the Serra d’Alto phase (fifth millennium BCE), is also

documented in the same cave by cinnabar (Aprile et al., 2017; Tiberi, 2019) found on a painted

brown cup in the Serra d’Alto style from the same Grotta dei Cervi cave. The only cinnabar

deposits mined in the Neolithic period are known in the Monte Amiata area (in Tuscany) and in the

eastern Adriatic area, mainly in Serbia (Šuplja Stena).

5. Conclusions

Molecular and isotopic investigation of archaeological materials excavated in Porto Badisco

and supposed to contain bitumen, have been presented. GC–MS-MS and EA–IRMS after

fractionation of the samples, permitted to identify bitumen on some archaeological potsherd

remains and to identify its geographic source by comparison with geological samples from Majella

and Selenicë, two deposits located respectively in Italy and Albania and known to have been

exploited since the Neolithic times. The data presented herein provide the first evidence of export

and trade of raw bitumen across the Adriatic Sea from Selenicë to Apulia during the Neolithic, thus

extending this transport across the Mediterranean to at least the fifth millennium BCE.
Acknowledgments

The authors would like to thank Dr. J. Connan and the other reviewers for their constructive and

helpfully suggestions.

Associate Editor–Bart van Dongen

References

Aprile, G., Bianco, S., Ingravallo, E., Muntoni, I.M., Tiberi, I., 2017. La Grotta dei Cervi di Porto

Badisco: gli scavi Lo Porto 1970–71. Rivista di Scienze Preistoriche 59–110.

Colombo, M., Tozzi, C., 2013. La cultura di Catignano nel panorama della ceramica dipinta

neolitica italiana: contatti con le culture della sponda adriatica orientale. In: Studies in

Mediterranean Archaeology for Mario Benzi. BAR International Series 2460, Oxford, pp.

249–259.

Connan, J., 1999. Use and trade of bitumen in antiquity and prehistory: molecular archaeology

reveals secrets of past civilizations. Philosophical Transactions of the Royal Society of

London. Series B: Biological Sciences 354, 33–50.

Connan, J., Dessort, D., 1989. Du bitume de la Mer Morte dans les baumes d’une momie

égyptienne. Comptes Rendus Academie Science de Paris 309 (II), 1665–1672.

Connan, J., Dessort, D., Aquitaine, E., 1987. Novel family of hexacyclic hopanoid alkanes. Organic

Geochemistry 11, 103–113.

Connan, J., Kozbe, G., Kavak, O., Zumberge, J., Imbus, K., 2013. The bituminous mixtures of

Kavuşan Höyük (SE Turkey) from the end of the 3rd millennium (2000 BC) to the Medieval

period (AD 14th century): Composition and origin. Organic Geochemistry 54, 2–18.

Connan, J., Nieuwenhuyse, O.P., Van As, A., Jacobs, L., 2004. Bitumen in early ceramic art:

bitumen-painted ceramics from Late Neolithic Tell Sabi Abyad (Syria). Archaeometry 46,
 115–124.

Connan, J., Nishiaki, Y., 2003. The bituminous mixtures of Tell Kosak Shamali on the Upper

Euphrates (Syria) from the Early Ubaid to the Post Ubaid: composition of mixtures and

origin of bitumen. In: Nishiaki, Y., Matsutani, T. (Eds.), Tell Kosak Shamali – The

Archaeological Investigations on the upper Euphrates Syria, Chalcholithic Technology and

Subsistence, vol. II, The University Museum and the University of Tokyo. Tokyo, pp. 283–

306.

De Pascale, A., Muntoni, I.M., 2018. Grotta dei Cervi di Porto Badisco. In: De Pascale, A.,

Tarantini, M. (Eds.), Segni dalla Preistoria. Siti dell’arte rupestre italiana nell’archivio

dell’Istituto Italiano di Preistoria e Protostoria. Istituto Italiano di Preistoria e Protostoria,

Firenze, pp. 206–233.

Faraco, M., Pennetta, A., Fico, D., Eramo, G., Beqiraj, E., Muntoni, I.M., De Benedetto, G.E.,

2016. Bitumen in potsherds from two Apulian Bronze Age settlements, Monopoli and Torre

Santa Sabina: Composition and origin. Organic Geochemistry 93, 22–31.

Freund, K.P., Batist, Z., 2014. Sardinian obsidian circulation and early maritime navigation in the

Neolithic as shown through social network analysis. The Journal of Island and Coastal

Archaeology 9, 364–380.

Graziosi, P., 1980. Le pitture preistoriche della grotta di Porto Badisco. Origines, Origines / a cura

dell’Istituto italiano di preistoria e protostoria. Giunti Martello.

Guglielmino, R., 2012. Il bitume di Roca. Breve nota su una sostanza negletta negli studi di

protostoria italiana. In: Annali della Scuola Normale Superiore di Pisa 4/2. pp. 99–114.

Ingravallo, E., Aprile, G., Tiberi, I., 2019. La Grotta dei Cervi e la preistoria nel Salento. Manni, p.

192.

Mattavelli, L., Novelli, L., 1990. Geochemistry and habitat of the oils in Italy. American

Association of Petroleum Geologists Bulletin 74, 1623–1639.

Nardella, F., Landi, N., Degano, I., Colombo, M., Serradimigni, M., Tozzi, C., Ribechini, E., 2019.
 Chemical investigations of bitumen from Neolithic archaeological excavations in Italy by

GC/MS combined with principal component analysis. Analytical Methods 11, 1449–1459.

Peters, K.E., Moldowan, J.M., 1991. Effects of source, thermal maturity, and biodegradation on the

distribution and isomerization of homohopanes in petroleum. Organic Geochemistry 17, 47–

61.

Peters, K.E., Moldowan, J.M., 1993. The Biomarker Guide: Interpreting Molecular fFssils in

Petroleum and Ancient Sediments. Prentice Hall.

Peters, K.E., Walters, C.C., Moldowan, J.M., 2005. The Biomarker Guide, Volume 1, Biomarkers

and Isotopes in the Environment and Human History. Cambridge University Press.

Rullkötter, J., Nissenbaum, A., 1988. Dead sea asphalt in egyptian mummies: Molecular evidence.

Naturwissenschaften 75, 618–621.

Schwartz, M., Hollander, D., 2008. Bulk stable carbon and deuterium isotope analyses of bitumen

artifacts from Hacinebi Tepe, Turkey: reconstructing broad economic patterns of the Uruk

expansion. Journal of Archaeological Science 35, 3144–3158.

Spataro, M., 2009. Cultural diversities: the Early Neolithic in the Adriatic Region and Central

Balkans. A pottery perspective. In: Gheorghiu, D. (Ed.), Early Farmers, Late Foragers, and

Ceramic Traditions: On the Beginning of Pottery in the Near East and Europe. Cambridge

Scholars Publishers p. 63.

Tiberi, I., 2019. Grotta dei Cervi a Porto Badisco. Un santuario per le genti del Mediterraneo (Scavi

Lo Porto 1970-71), Millenni. ed. Museo e Istituto Fiorentino di Preistoria “Paolo Graziosi,”

Firenze.

van der Werf, I.D., Fico, D., De Benedetto, G.E., Sabbatini, L., 2016. The molecular composition of

Sicilian amber. Microchemistry Journal 125, 85–96.

Zilhão, J., 2014. Early prehistoric navigation in the Western Mediterranean: Implications for the

Neolithic transition in Iberia and the Maghreb. Eurasian Prehistory. Island Archaeology and

the Origins of Seafaring in the Eastern Mediterranean 11, 185–200.

Figure captions

Fig. 1. Pictures showing two shards from which bitumen was scraped and analysed.

Fig. 2. Gas chromatography–mass spectrometry (GC–MS) analysis of the saturated hydrocarbon

fraction of PB2 sample from Porto Badisco. The total ion current shows a moderate degree of

biodegradation.

Fig. 3. C27 and C29–35 hopane distribution for PB2–PB4 archaeological samples along with an

Albanian sample (SE2). GC–MS-MS parent–daughter transition data are reported on the top of each

column.

Fig. 4. C24 tetracyclic terpane/C23 tricyclic terpane vs gammacerane/C29 17α(H),21β(H)-hopanes for

archaeological and geological samples.

Fig. 5. GC–MS-MS ion chromatograms of extractable C27–C30 steranes from Porto Badisco

archaeological samples along with an Albanian sample (SE2). The relative signal intensity of each

sterane channel shows a C29 dominance. The parent–daughter transitions are reported on the top of

each column.

Fig. 6. Plot of the δ13C vs δ34S isotopic ratio of asphaltene fractions.

Tables

Table 1. Description of the archaeological sample from Grotta dei Cervi at Porto Badisco and their

relevant gross properties

Sample Archaeological Description Mass EOM EOM Asphaltene

reference
g g % %
PB1 ex114 body sherd 0.060 0.002 3
PB2 ex153 handle of pottery in 0.103 0.029 28 24
the Serra d’Alto style
PB3 ex412 bottom of a vessel 0.055 0.014 25 20
PB4 ex208 body sherd 0.036 0.011 31 6
PB5 ex414 handle of pottery in 0.015 0.001 7
the Serra d’Alto style
Table 2. Selected biomarkers for archaeological and geological samples that have been used as

source parameters in the present paper. Tricyclic and tetracyclic terpanes were determined by GC–

MS whereas pentacyclic terpanes and steranes by GC–MS-MS (for transitions see Faraco et al.,

2016).

Label SE1 SE2 SE3 MJ1 MJ2 MJ3 PB2 PB3 PB4
T24/T23 0.33 0.33 0.38 0.29 0.36 0.31 0.32 0.45 0.33
Tetra24/T23 1.17 1.01 1.64 2.00 2.04 2.25 1.01 1.14 1.35
Ts/Tm 0.09 0.10 0.15 0.18 0.15 0.21 0.12 0.14 0.10
H29/M29 17.3 19.1 3.11 17.3 16.9 18.0 20.2 24.7 20.7
H29/H30 1.27 1.22 15.5 4.20 1.61 20.9 1.35 1.51 1.77
H30/M30 42.2 33.8 1.63 8.74 25.2 1.71 35.0 40.5 31.8
G/H29 0.02 0.03 0.32 0.27 0.20 0.10 0.03 0.04 0.02
H31 41% 43% 20% 35% 50.5% 39% 42% 41% 44%
H32 22% 23% 23% 23% 21.1% 22% 21% 22% 23%
H33 18% 16% 19% 20% 10.6% 16% 17% 17% 16%
H34 10% 10% 14% 12% 11.2% 12% 10% 11% 10%
H35 8% 9% 23% 10% 6.6% 12% 9% 9% 9%
IHR33 0.04 0.07 1.13 0.16 0.08 0.05 0.05 0.03 0.04
IHR34 0.07 0.05 1.54 0.14 0.08 0.08 0.06 0.05 0.06
N33 0.11 0.11 4.19 0.51 0.25 0.21 0.11 0.10 0.10
N34 0.15 0.17 5.25 0.57 0.13 0.24 0.14 0.15 0.11
2C35/6/(H35
 R+S) 0.11 0.20 3.09 0.52 0.68 0.34 0.10 0.14 0.14
2C34/6/(H34
 R+S) 0.14 0.37 7.33 0.86 0.88 0.80 0.17 0.14 0.17
27S 26% 23% 19% 60% 29% 58% 28% 22% 23%
28S 21% 15% 35% 10% 17% 9% 12% 15% 13%
29S 54% 62% 46% 31% 54% 33% 60% 63% 64%
29S/(S+R) 0.55 0.56 0.40 0.67 0.43 0.31 0.50 0.57 0.54
D/S 0.47 0.56 6.45 0.07 0.03 0.11 0.49 0.91 0.55
T24/T23: C24 tricyclic terpane/C23 tricyclic terpane; Tetra24/T23: C24 tetracyclic terpane/C23

tricyclic terpane; Ts/Tm: 18α-22,29,30-trisnorneohopane/22,29,30-trisnorhopane; G/H29:

gammacerane/17α(H),21β(H)-30-norhopane; H29/H30: 7α(H),21β(H)-30-

norhopane/17α(H),21β(H)-hopane; H: 17α(H),21β(H)-hopanes; H29/M29: 17α(H),21β(H)-30-

norhopane/17β(H),21α(H)-30-norhopane; H30/M30: 17α(H),21β(H)-hopane/17β(H),21α(H)-

hopane; H31,…, H35: % of total C31–C35 -homohopanes; IHR33: C33 isohopanes/regular C33

hopanes; IHR34: C34 isohopanes/regular C34 hopanes; N33: C33 30-norhopanes/regular C33 hopanes;

N34: C34 30-norhopanes/regular C34 hopanes; 2C34/6/(H34  R+S): 2C34

hexahydrobenzohopane/C34 -homohopanes (Connan et al., 1987); 2C35/6/(H35  R+S): 2C35

hexahydrobenzohopane/C35 -homohopanes (Connan et al., 1987); 27S: %C27 steranes/(C27 + C28

+ C29 steranes); 28S: %C28 steranes/(C27 + C28 + C29 steranes); 29S: %C29 steranes/(C27 + C28 + C29

steranes); 29S/(S+R): C29 steranes, 20S/(20S + 20R); D/S: C27–29 diasteranes/C27–29 regular steranes.
Table 3. Carbon and sulfur elemental analyses and isotopic fingerprints relevant to asphaltene

fraction of Porto Badisco, Selenicë and Majella samples (m ± 1 sd).

C S δ13C δ34S
Sample
% % ‰, vs VPDB ‰, vs VCDT

PB2 71.5 ± 1.8 7.6 ± 0.4 –29.26 ± 0.07 –23.9 ± 0.1

PB3 67.3 ± 1.6 7.5 ± 0.5 –29.32 ± 0.04 –24.2 ± 0.1

PB4 71.7 ± 1.2 7.7 ± 0.4 –29.12 ± 0.17 –23.2 ± 0.2

MA1 74.3 ± 1.1 11.5 ± 0.5 –28.68 ± 0.03 –20.9 ± 0.1

MA2 68.1 ± 1.7 10.2 ± 0.4 –28.01 ± 0.07 –20.2 ± 0.1

MA3 74.1 ± 2.5 10.9 ± 0.2 –28.49 ± 0.16 –19.4 ± 1.5

SE1 73.7 ± 2.0 8.7 ± 0.6 –29.26 ± 0.10 –23.8 ± 0.3

SE2 74.9 ± 0.2 9.5 ± 0.7 –29.11 ± 0.05 –23.1 ± 0.2

SE3 74.2 ± 0.6 8.3 ± 0.1 –29.01 ± 0.10 –23.4 ± 0.3

Declaration of interests

☒ The authors declare that they have no known competing financial interests or personal
relationships that could have appeared to influence the work reported in this paper.

☐The authors declare the following financial interests/personal relationships which may be considered as
potential competing interests:
Highlights

 First geochemical study of bitumens from Porto Badisco (Italy) archaeological site
 Albanian origin of the bitumens found on Neolithic ceramics from Porto Badisco
 First evidence of bitumen trade across the Adriatic Sea from the 5th millennium BCE