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XIII/1/2022
INTERDISCIPLINARIA ARCHAEOLOGICA
NATURAL SCIENCES IN ARCHAEOLOGY
homepage: http://www.iansa.eu
A look at the region
Five Years of Advanced Archaeometric Analysis at the Department of
Analytical Chemistry, Faculty of Science, Palacký University in Olomouc
Lukáš Kučera
1*
, Petr Bednář
1
1
Department of Analytical Chemistry, Faculty of Science, Palacký University in Olomouc, 17. listopadu 1192/12, 771 46, Olomouc, Czech Republic
1. Introduction
Organic residues in archaeological fndings are mostly
characterised by archaeobotanical and archaeozoological
expertise and common microscopic methods. For their
chemical characterisation the focus is usually on the
determination of several basic parameters,
e.g.
, total organic
carbon, total nitrogen, and the content of phosphate, sulphur,
sodium, potassium, calcium, and magnesium. However,
a more systematic chemical analysis of organic residues in
an archaeological context using modern analytical techniques
is often lacking (Gofer, 2005).
The residues of colorants and pigments on the surface
of ancient material are studied using various procedures,
as described in several articles (Leon, 2012; Pozzi, 2012;
Parras, 2010). The most frequent techniques are based on
mass spectrometry with various ionization techniques (
i.e.
,
MALDI, ICP-MS, and ESI) and microscopic techniques (
i.e.
,
PIXE, Raman microspectrometry, SEM, and EDX). Some of
these methods are commonly used in archaeology, mainly
for non-organic analysis, for example, mineral composition
of prehistoric ceramics (Santos Rodrigues, 2015) or metal
artefacts (Oudbashi and Shekofteh, 2015). On the other hand,
much less attention has been paid to an objective analysis of
food residues in ancient fndings, although these materials
when studied comprehensively ofer opportunities to obtain
unique information. In many interesting cases, food residues
have been found in localities connected with obsequies and
other ancient mysteries. Chemical analysis can also help to
prove some historical hypotheses, for example, the presence
of garlic in graves as anti-vampire prevention and the use
of various plant drugs (
Cannabis sativa L.
, poppy extract,
opium,
etc.
) in rituals (Askitopoulou, 2002). Perhaps the
most inspirational approach well-worth following was called
the Archaeological Biomarker Concept, as introduced by
Evershed (2008).
In an efort to systematise analytical research in the feld of
analysis of organic and composite residues in archaeological
fnds, a project entitled “Advanced chemical analysis of
residues of organic materials in archaeological context”
was prepared by our team at the Department of Analytical
Chemistry, Faculty of Science, Palacký University in
Olomouc in cooperation with the Archaeological Centre,
Olomouc, in 2016 and received funding the following year
Volume XIII ● Issue 1/2022 ● Pages 85–90
*Corresponding author. E-mail: lukas.kucera@upol.cz
ARTICLE INFO
Article history:
Received: 10
th
January 2022
Accepted: 10
th
February 2022
DOI: http://dx.doi.org/10.24916/iansa.2022.1.7
Key words:
archaeometry
analytical chemistry
organic residues
spectrometry
spectroscopy
chromatography
ABSTRACT
In modern archaeological research, a close multidisciplinary collaboration with other scientifc areas
is necessary, especially with natural sciences (
e.g.
, anthropology, archaeobotany, and chemistry). This
kind of collaboration and mutual evaluation of obtained results provides synergistically a series of
important information in the context of prehistoric research nowadays. This systematic cooperation
among archaeology, heritage science, anthropology, archaeobotany and analytical chemistry has been
intensively developed for last fve years at the Department of Analytical Chemistry, Faculty of Science,
Palacký University in Olomouc. The aim of this short communication is to introduce our workplace
and its activities with a focus on the most important outputs from various areas of the archaeometric
research.
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IANSA 2022 ● XIII/1 ● 85–90
Lukáš Kučera, Petr Bednář: Five Years of Advanced Archaeometric Analysis at the Department of Analytical Chemistry, Faculty of Science,
Palacký University in Olomouc
86
from the Czech Science Foundation. This project helped to
establish a wider group of cooperating experts from various
areas and institutions: the Laboratory of Archaeobotany
and Paleoecology (LAPE), Laboratory of Morphology
and Forensic Anthropology (LaMorFA), Institute for
Archaeological Heritage (UAPP), National Heritage
Institute (NPU), Institute of Archaeology of the Czech
Academy of Sciences in Prague (ARÚP AV ČR), the Czech
Numismatic Society (mainly the Prague Groschen branch
ofce) and other specialised workplaces. Cooperation
has also been established at an international level, for
example with the University of Wroclav (Poland), Eastern
Mediterranean University (N. Cyprus), National Museum
in Belgrade (Serbia), Klaipeda University (Lithuania)
and the Italian Institute for Conservation and Restoration
under the Ministry of Culture (ISCR, Italy). Subsequently,
a project in the Operational Programme of the European
Union entitled: “Advanced physical-chemical methods of
research and protection of cultural and artistic heritage”
(OA ITI – ARTECA) was obtained and expanded the
expertise of our laboratory to include historical art. The
advanced analytical research is performed using modern
scientifc instrumentation, including six mass spectrometers
with various mass analysers (quadrupole, triple quadrupole,
Figure 1.
A: Photo of the bronze situla (“bucket”) of Kurd type, Kladina variant (photo Luděk Vojtěchovský); B: Chromatograms of MRM transition 440 →
189 of soil extracts and miliacin standard.
Figure 2.
A: Part of the ceramic pan KE2; B: GC/MS analysis of acetone:chloroforme extract of samples – detection of cholesterol (samples of organic
residue from the pit, KE2-1 and KE2-3; samples KE2-2 and KE2-4 were taken under KE2-1 and KE2-3, respectively; reference sample from the bottom of
the pan, KE2-5; inner surface KE2-6 and reference sample from the surface, KE2-7).
image/svg+xml
IANSA 2022 ● XIII/1 ● 85–90
Lukáš Kučera, Petr Bednář: Five Years of Advanced Archaeometric Analysis at the Department of Analytical Chemistry, Faculty of Science,
Palacký University in Olomouc
87
ion-trap, and hybrid quadrupole-time of fight) and various
ion sources (electron impact and chemical ionisation,
electrospray, chemical ionisation and photoionisation at
atmospheric pressure, atmospheric solids analysis probe,
desorption electrospray, and vacuum laser desorption-
ionisation (MALDI source)). These mass spectrometers are
routinely hyphenated with separation techniques,
i.e.
, GC,
UHPLC and 2D-UHPLC as well. Two mass spectrometers
(Synapt G2-S and Cyclic IMS) are equipped with an ion
mobility cell allowing the separation of ions in accordance
with their collision cross sections. Note that cyclic ion
mobility is a unique and extremely efcient approach to
separate compounds based on their collision cross sections,
including very closed structures and true isomers (the
recent installation of a Cyclic IMS system in the applicants’
laboratory was only the third one in the world). The obtained
results are continuously published in prestigious scientifc
journals. In the following text, the most important results
recently obtained by our team are briefy reviewed.
2. Case studies
2.1 Organic residues of food and beverages
The soil content of two ceramic vessels belonging to the
Moravian regional group of the Corded Ware Culture was
analysed using the MALDI-MS technique. Several markers
found in the soil layers taken from the bottom of the studied
vessels correspond with signals of triacylglycerols typical
for milk and/or dairy products. These results represent the
frst direct evidence of the utilisation of milk products in
the Eneolithic period in the Moravian Corded Ware Culture.
Moreover, triterpenoid miliacin that is a marker of millet
was detected in this soil material by atmospheric solids
analysis probe mass spectrometry (ASAP-MS) and gas
chromatography/mass spectrometry (GC/MS). It should
be emphasised that this was the frst time the ASAP-MS
technique was used in archaeological research worldwide.
The presence of millet in an Eneolithic context is very
important and extraordinary. It can be considered as the frst
such direct evidence of millet usage in Central Europe. Our
data relating to the utilisation of milk and millet in the Early
Neolithic–Eneolithic period in Eastern Central Europe (the
Central Danube Region) signifcantly extend the former
evidence of dairy products and millet usage in Western
Central Europe (Germany, Switzerland) (Kučera
et al.
,
2018; Kučera
et al.
, 2019). The miliacin was also detected
on the surface of a bronze situla from the Kladina site in East
Bohemia (Pardubice district). The detection of miliacin (as a
“chemical imprint” of millet; Figure 1) by chemical analysis
opens the way for a better understanding of the composition
of food remains, even in much older samples than those
studied up to now, or procedures that would lead to complete
corn decomposition (
e.g.
, cooking, baking, fermentation,
etc.
). Archaeobotanical analysis (pollen and starch grains)
pointed to the use of bitter herbs in a fermentation process. It
can be concluded that this bronze situla contained the oldest
(yet) millet-herbal beer in Europe (Jílek
et al.
, 2021). The
presence of millet in studied archaeological material has also
been described in the following publications (Kučera
et al.
,
2020; Golec
et al.
, 2022). As can be seen, the combination
of archaeobotanical and chemical analyses of archaeological
samples is a powerful tool for revealing new information
from human history. The next example is the analysis of
Neolithic ceramic fragments of large, fat, elongated pans
from North Macedonia (Figure 2). The attached material
on the surface of the studied pan fragments was sampled
for consequent chemical and microscopical analyses (
i.e.
,
analyses of starch, phytoliths, and microscopic animal
remains). The main compound that occurred in all samples
was cholesterol (found using GC/MS). A major source of
cholesterol is animal fat and meat (its presence being proved
by an immunological test). Based on these results, we
suppose that the analysed ceramic pans from Ustie na Drim
were used for the preparation of meals containing meat from
common livestock in combination with cereals and wild
plants (Beneš
et al.
, 2021).
2.2 Metal analyses
Another area of current research is dealing with the analysis
of metallic objects and the presence of organic residues on
their surface (coins, ritual objects, decorations, jewels,
etc.
).
The composition of a metal surface can infuence the organic
residues that adhere to its surface (for example, catalytic
action, formation of organometallic adducts, etc) and, on
the other hand, organic material can cause changes in the
metal surface itself (for example, corrosion protection, either
intended or accidental). A detailed analysis of coins from
the Ladná hoard (a fnding of more than 1000 coins; broad
pfennigs and Moravian denarii discovered in the 1990s) was
performed. Several methods were applied for assessing the
original and actual fneness of the broad silver pfennigs from
this hoard (Kučera
et al.
, 2017). The knowledge from the
above studies dealing with coin characterisation was applied
in a study of the defects found on historical gold coins known
as “gold corrosion” (Figure 3). Defects analysed in this study
show a high content of iron and oxygen. The results show
that silver and the consequent formation of Ag
2
S is not the
only possibility for the formation of red stains. The coins that
are minted in thousands/millions of pieces have a higher risk
of damaging the parts of devices used in the mint and thus
contaminating a golden alloy with iron particles. This study
expands our knowledge of the formation of “corrosion”
products on gold coin surfaces and can be the primary impulse
to control steel components during the minting process
(Kučera
et al.
, 2021). Another study dealt with the purpose
of use of a bronze ornithomorphic plastic artwork, coming
from an extensive hoard from the territory of the Liptov Basin
(Central Slovakia), belonging, in terms of art, to some of the
crucial artefacts of the European Bronze Age. GC/MS revealed
the presence of particular fatty acids (isomyristic, oleic,
stearic), monoacylglycerols (monomyristin, monopalmitin,
monostearin) and cholesterol. Raman microscopy proved
the presence of soot (based on signals at 1323 and 1589 cm).
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IANSA 2022 ● XIII/1 ● 85–90
Lukáš Kučera, Petr Bednář: Five Years of Advanced Archaeometric Analysis at the Department of Analytical Chemistry, Faculty of Science,
Palacký University in Olomouc
88
X-ray fuorescence spectrometry confrmed that the material
of the artwork is bronze. It can be concluded on the obtained
data and formerly published studies that this object was used
as a ceremonial lamp (Ondrkál
et al.
, 2020). These studies
particularly highlight the importance of the careful, but critical,
interconnection of data that have been obtained from various
analytical techniques and with archaeological expertise.
Recently, three copper alloy moulds from Moravia
belonging to the Roman era were analysed by several analytical
techniques, including XRF, GC/MS and a combination of
atmospheric solid analysis probe-mass spectrometry with
ion mobility spectrometry. The elemental composition (
i.e.
,
the higher content of Pb and Zn) of two moulds pointed
to the possibility of the production of “barbarian” alloys
from remelted Roman objects. The internal part of all
moulds contained carbon in the form of soot (identifed by
Raman microscopy). Moreover, the moulds contained solid
material that was identifed as a residue of ozokerite (earth
wax, a naturally-occuring odoriferous mineral wax). We
hypothesise that the remains of earth wax in these moulds
point to their use for the production of wax models and the
consequent use of these models for lost‐wax ceramic casting
(Jagošová
et al.
, 2021).
2.3 Exploration of works of art
The XRF, a common and fast technique for metal analysis,
can be advantageously used for the non-invasive exploration
of works of art, for example, pigments and plasters. This
kind of application was performed by our team in the case
of a painting in The Tombs of the Via Ostiense in Rome
(Figure 4; Marcelli
et al.
, 2020).
An investigation of a scapular amulet from the collection
of the Regional Museum in Olomouc was also conducted by
a combination of various methods. GC/MS of four micro-
samples taken revealed the presence of dehydroabietic acid.
The LDI-MS technique proved the presence of fatty acids
as well as di- and triacylglycerols. Common organic dyes or
pigments were not detected. The dark colour of the material
is thus more related to inorganic black pigments (
e.g.
, soot)
than anything else. The high content of lead was found by
Figure 3.
A: Austro-Hungarian 10 Korona 1905 afected by corrosion; B: Raman imaging of corrosion products on surface of Austro-Hungarian 10 Korona
at the signal 394 cm
−1
(the lowest intensity of Raman signal is shown by the blue colour and the highest intensity by the red colour); C: Raman spectra of 10
Korona and reference materials – area of pure gold; D: Analysis of red stain with laser energy 10 mW; E: Analysis of red stain with laser energy 20 mW; F:
Spectrum of hematite standard; G: Spectrum of goethite standard.
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IANSA 2022 ● XIII/1 ● 85–90
Lukáš Kučera, Petr Bednář: Five Years of Advanced Archaeometric Analysis at the Department of Analytical Chemistry, Faculty of Science,
Palacký University in Olomouc
89
ICP-MS, which was ascribed to the presence of siccative.
With the support of these results, the studied material was
described as an oil paint containing desiccant (Janusová
et al.
, 2017).
2.4 Analyses of resins and related compounds
The combination of untargeted LDI-MS with MVA was
applied to the classifcation of some amber fndings. The
composition of amber varies according to its origin. However,
diferences in its composition are refected by relatively
small changes in a huge number of signals that are nearly
impossible to evaluate by (manual) spectral comparison
and the classifcation of the ambers according to place of
origin (geographical distribution) – and thus this task is
predestined for MVA analysis. Amber samples from various
archaeological sites of Western Lithuania and its coast were
studied. The total segregation of raw amber samples from
a strip-mine (Palmnicken in former East Prussia in the
Sambian Peninsula, currently Kaliningrad Oblast, Russian
Federation) and the seacoast (Lithuania) was observed in
a Score Plot from a PCA analysis. The distribution of samples
with respect to diferent localities was observed mainly in
the direction of the frst component axis (explaining 59.2%
of data variation; the frst two principal component together
explaining 82.2% of the variation) (Bliujiene and Kučera,
2018). This technique was used for the characterisation of
amber fndings from the basal layers at the Pod Hradem
Cave (Moravian Karst) (Nejman
et al.
, 2018) and amber
samples from Mikulovice (Kučera and Bednář, 2020). The
majority of amber is found in graves. In addition, other
hardened resins (or some other natural compound) could
be found. One example from the grave of an older woman
attributed to the Iron Age Horákov Culture was the residue
of hardened birch bark tar (identifed by GC/MS), possibly
a form of natural medicine (Holubová
et al.
, 2020).
2.5 Analyses of anthropological samples
An interesting task was to analyse human teeth from some
archaeological excavation to fnd a link between phosphate
intensity changes in the human enamel and the light
microscopy record of accentuated stress lines (ALs). This
principle of dental “time-lapse records” is increasingly
used for analytical purposes in palaeoanthropology and
bioarcheology to reconstruct individual life histories
from archaeological human skeletal remains. This human
“barcode” pattern is unique to each person, preserving
a record of individual life experience. Based on the
obtained results, we suppose that Raman microscopy of
ALs could ofer a new and alternative perspective on
enamel structures. In most cases, we found that the optical
change in the light microscope corresponds to the change
in phosphate concentration in the Raman microscope.
However, Raman microscopy may not detect everything
that is recognised by light microscopy. Raman microscopy
can help us to decide which changes in the stress lines are
related to changes in phosphate content and which are not
(Vacková
et al.
, 2021).
3. Conclusion
Our fve years of experience in the feld have shown
conclusively that the very close cooperation of an analytical
chemist with an archaeologist/art expert represents the
principal premise for a reliable and coherent research of
historical samples – from the disclosure of an archaeological
locality and sample collection through to the analysis and
data interpretation.
This close cooperation of analytical chemistry with
archaeological area is refected in our recent book (Bednář
and Kučera, 2021). The book is devoted to the utilisation
Figure 4.
A: Tomb VII at the Via Ostiense in Rome; B: Place of sampling and XRF measurement; C: Detail of sample for consecutive preparation of cross-
section; D: Stratigraphy of studied sample – the blue layer contained 33 % copper,
i.e.
, blue copper pigment.
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IANSA 2022 ● XIII/1 ● 85–90
Lukáš Kučera, Petr Bednář: Five Years of Advanced Archaeometric Analysis at the Department of Analytical Chemistry, Faculty of Science,
Palacký University in Olomouc
90
of modern techniques and methods of chemical analysis in
the research of human history. It is intended for historians to
provide them with detailed information on the possibilities,
as well as the pitfalls, of current analytical procedures and
for analytic chemists interested in research in the feld of
history, archaeology and cultural heritage. The book is freely
available at www.vydavatelstviupol.cz as an iPDF.
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