Microchemical Journal 125 (2016) 1020

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Microchemical Journal

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Microanalysis of clay-based pigments in paintings by XRD techniques

David Hradil a,b,, Petr Bezdika a,b, Janka Hradilov b, Vlasta Vautov b
a
Institute of Inorganic Chemistry of the Czech Academy of Sciences, v.v.i., ALMA Laboratory, 1001 Husinec-e, 250 68 e, Czech Republic
b
Academy of Fine Arts in Prague, ALMA Laboratory, U Akademie 4, 170 22 Prague 7, Czech Republic

a r t i c l e i n f o a b s t r a c t

Article history: Despite considerable advantages of the method in the eld of microanalysis of pigments, powder X-ray micro-
Received 15 July 2015 diffraction has not yet been satisfactorily tested for correct and unambiguous discrimination of clay structures
Received in revised form 15 October 2015 in colour layers of paintings and, eventually, for reliable quantitative analysis of their relative contents. To over-
Accepted 24 October 2015
come this lack of knowledge, several sets of laboratory experiments with model mixtures and reference samples
Available online 30 October 2015
imitating real paint layers have been performed and general rules for proper discrimination of clay structures
Keywords:
have been described. As an integral part of the research, claybinder interactions were studied to avoid mislead-
Clay minerals ing interpretations. It was found out that various characteristic shifts of basal diffraction lines depend on the type
Earth pigments of organic binder and the relative content of expandable layers in the clay structure. Based on these results, it was
Paintings possible to differentiate pure smectites, interstratied illite/smectite structures and chlorites in paint and prepa-
Powder X-ray micro-diffraction ratory layers of actual paintings. The accuracy of quantitative microanalysis of clay minerals in mixtures with
Quantitative phase microanalysis other minerals in Mid-European red earth pigments (typically containing kaolinite, illite, quartz, hematite, and
Claybinder interaction anatase) was characterised by total bias values in the range of 310 wt.%. Finally, results were applied to differ-
entiation of earth pigments of various provenances in grounds of Baroque paintings and Early Renaissance prep-
aration layers for gilding.
2015 Elsevier B.V. All rights reserved.

1. Introduction relatively low Raman scattering of clay structures, particularly in mix-

In ancient times, clay minerals represented a substantial part of
earth colours in the rock as well as applied art. Later on, they were
employed in panel and easel paintings not only as pigments, but also
as llers, grounds, and preparatory layers for gilding [1]. In the painting
technology, the content of clay minerals is appreciated for the increase
of adhesive power of the material; on the other hand, higher amounts
of expandable clay structures may lead to an increased risk of swelling
under wet conditions. Analytical characterisation is essential for the ex-
planation of relations between clay minerals' composition and material
properties, but it is also important for provenance studies, as clays are
sensitive to physicochemical conditions of the process of their natural
formation. A number of standard analytical procedures commonly
applied to clays (preparation of oriented specimen, ethylene glycol
solvation etc.) [2] cannot be used when dealing with minute amounts
of heterogeneous materials obtained from cultural heritage objects. In
addition, Raman micro-spectroscopy, being otherwise suitable for
mineralogical microanalysis, cannot be efciently applied due to the
Selected papers presented at TECHNART 2015 Conference, Catania (Italy), April
2730, 2015.
Corresponding author at: Institute of Inorganic Chemistry of the Czech Academy of
Sciences, v.v.i., ALMA Laboratory, 1001 Husinec-e, 250 68 e, Czech Republic.
E-mail addresses: hradil@iic.cas.cz (D. Hradil), petrb@iic.cas.cz (P. Bezdika),
hradilovaj@volny.cz (J. Hradilov), vasutovavlasta@seznam.cz (V. Vautov).
tures with iron and titanium oxides [3]. A missing methodology is
thus the main reason why clay minerals are not being properly detected
and differentiated in colour and preparatory layers of paintings regard-
less of their often signicant contents in pigment mixtures.
As the sampling of historical paintings is restricted and the obtained
micro-samples are rare, a non-destructive analytical approach prevails.
Conventionally, paint micro-samples (typically at paint chips with di-
ameter not exceeding in any direction 1 mm) are embedded in synthet-
ic resin and polished to obtain cross-section. These micro-sections are
considered to be the standard samples, as the stratigraphy of paint
layers is visualised and further analyses can be performed on individual
layers. In rare cases, the material obtained from the painting is large
enough to be split, and one of the fragments can be analysed separately
without any pre-treatment. Routinely, description of the sample by the
means of optical microscopy is followed by semi-quantitative elemental
analysis (e.g., by scanning electron microscopy coupled with energy-
dispersive spectroscopy SEM/EDS or micro X-ray uorescence
micro-XRF), and by spectroscopic analyses of pigments and binders by
Raman and Fourier-transform infrared micro-spectroscopy (micro-
Raman, micro-FTIR). None of these methods leads to the differentiation
of clay minerals due to their variable chemical composition further
affected by numerous admixtures, structural similarities, and low
Raman scattering. To identify individual clay minerals, it is necessary
to apply powder X-ray micro-diffraction (micro-pXRD), which repre-
sents still a rather unconventional approach in the eld of paint analysis.

http://dx.doi.org/10.1016/j.microc.2015.10.032
0026-265X/ 2015 Elsevier B.V. All rights reserved.
 D. Hradil et al. / Microchemical Journal 125 (2016) 1020
Synchrotron-based micro-pXRD has been used, for example, to identify
pigments in Roman paintings [4], to quantify mineral components in
the natural lapis lazuli (ultramarine) blue pigment found in the 15th
century altarpiece [5], or to detect different crystal forms of the lead
chromate yellow when explaining its degradation of in paintings by
Vincent van Gogh [6]. A combination of synchrotron- and laboratory-
based micro-pXRD techniques have been applied to characterise pig-
ments in Pompeian wall paintings [7] or in Baroque paintings by Span-
ish painter Murillo (16171682)clay minerals were identied in the
ground layer, but were not further differentiated [8]. Nevertheless, in
one of our previous articles, we have demonstrated that clay minerals'
structures in old paints can be characterised in detail (e.g., by specic
parameters like crystallinity indices), and that the results of synchrotron
and laboratory micro-pXRD were highly comparable [9]. On the other
hand, the identication of clay minerals by portable XRD techniques
[10,11] is prevented by technical limitations of goniometers which do
not reach low angles, where most of the characteristic diffraction lines
of clay minerals do appear. One of the potentially perspective tech-
niques overpassing this problem is energy-dispersive XRD, which is
now appearing also in portable conguration for in situ measurements
[12].
One of the main limits of laboratory micro-pXRD is the size of the ir-
radiated area. A mono-capillary with an exit diameter of, e.g., 0.1 mm,
creates an irradiated area delimited by an ellipse, whose smaller diam-
eter (perpendicular to the beam) is ca. 0.140 mm and larger diameter
(parallel with the beam) ranges between ca. 4 and 0.22 mm for 2
theta angles of 480 [13]. Therefore, in low angles, it can exceed the
size of the paint layer of interest or even the size of the whole micro-
sample of the painting. Consequently, thicker layers are easier to
study, such as, e.g. the clay-based grounds, while individual grains of
earths present in certain paint layers are too small to be analysed [14].
Another limitation of the micro-pXRD setup is that the beam of the
mono-capillary is only quasi-parallel and has an instrumental width
(FWHM, full width in half maxima) of about 0.3. Therefore, micro-
pXRD has worse resolution than the conventional BraggBrentano ge-
ometry and, consequently, the limits of reliability of quantitative
phase analysis (QPA) based on micro-pXRD and corresponding errors
in QPA are higher, rising up to about 5% (w/w). Finally, the last substan-
tial limitation that inuences the micro-pXRD analysis is the preferred
orientation of grains (typical for at clay particles), which cannot be
avoided in solid samples, represented in this study by fragments and/
or cross-sections obtained from paintings. These samples have to be
analysed in a non-destructive way and remain preserved for other ana-
lytical techniques or archiving. Therefore, the preferred orientation can-
not be prevented.
There is still a substantial lack of knowledge corresponding with the
fact that micro-pXRD technique has not yet been satisfactorily tested for
correct and unambiguous discrimination of clay structures in paint
layers and, eventually, for reliable semi-quantitative analysis of their
relative contents in the mixture. Furthermore, the interpretation of
clay structures in terms of technological or art-historical contexts is
still an unexplored eld. As a result, clay minerals in paint layers are rou-
tinely being described only by general terms (such as, e.g., clays or
aluminosilicates), or they are completely omitted in the materials re-
search of paints. Terms like ochres or earths are associated more likely
with the presence of iron oxides affecting their colour despite the vari-
able contents of clay minerals ranging from 0 to almost 100 wt.%. These
variable contents reect the natural process of formation and, conse-
quently, also the provenance of the material, and strongly affect its tech-
nological properties.
Clay structures can be divided into two major groups according to
their ability to expandexpandable and not-expandable. Expand-
ability refers to the ability of the structure to extend its interlayer
space by adsorbing large polar molecules from the surrounding envi-
ronment. The positions of basal diffraction lines directly indicate the
presence of most of the non-expandable structures (such as, e.g., the
11
most frequent kaolinite at d001 = 0.710.72 nm or clay micaillite at
approx. d001 = 1.00 nm). However, the most problematic is the differ-
entiation of expandable structures, which exhibit their 001 diffraction
lines at various positions in very low angles (d001 N 1.00 nm). The
actual basal spacing corresponds to (i) the type of the mineral
(e.g., Mg-vermiculite at approx. 1.44 nm and Mg smectite at approx.
1.50 nm), (ii) the type of the interlayer cations, and the state of hydra-
tion (e.g., Ca/Mg smectite at approx. 1.50 nm, but Na smectite at approx.
1.30 nm), and nally, (iii) the presence of interstratied structures,
i.e., mixed crystals containing both expandable and non-expandable
layers, where the actual position reects the relative ratio of these two
types of layers and varies, for example, from 1.00 to 1.50 nm in the case
of randomly ordered interstratied illitesmectite structures (the most
frequent clay mixed crystal in nature). In addition, it is not easy to differ-
entiate expandable clay minerals from the group of non-expandable clay
chlorites with basal spacing at approx. d001 = 1.42 nm. To unambiguously
identify these structures, powdered clay material is usually intercalated
by specic organic molecules (such as ethylene glycol) in order to gener-
ate highly characteristic changes in the diffraction pattern of expandable
phases [2]. However, such procedure cannot be performed on painting
micro-samples or their cross-sections. Further, it is very probable, and
yet has never been studied, that expandable clay structures in paints
and paint layers are affected by interaction with organic binders. These in-
teractions may hamper the identication of the original structure, which
is modied into an organo-clay intercalate. In the classical literature, but
also in recent studies, there are examples of intercalation of clays by
chemical compounds, which could also appear in historical paint binders
(e.g. fatty acids or peptides) [1519]. Nevertheless, results of none of
these studies, which are primarily directed to chemicaltechnological, ag-
ricultural, or industrial applications, cannot be directly applied to complex
mixtures of historical paints, which signicantly differ in composition and
physical properties.
Regarding the current knowledge, the main objective of this study
was to nd general rules for proper discrimination of clay structures
in either colour or preparatory layers of paintings using micro-pXRD
technique. This included experimental study of claybinder interactions
and the testing of limits, accuracy, and reliability of quantitative phase
analysis of complex mixtures on model and actual paintings' samples.
The study of claybinder interactions was motivated by numerous nd-
ings of diffraction lines corresponding to basal spacing exceeding
1.20 nm in actual ground layers of paintings, which indicated the possi-
ble presence of expandable clay minerals. Due to the great variability of
interstratied structures with variable contents of expandable layers in
nature, it was necessary to know to what extent the position of their
characteristic diffraction lines is inuenced by the interaction with the
binderprior to the nal interpretation of the structure. The second
task, focused on the quantitative microanalysis, is particularly impor-
tant for clays formed in temperate climate (such as, e.g., Central
Europe) that frequently contain the same major phases (typically kao-
linite, clay micaillite, quartz, hematite or goethite, and anatase), but
in different proportions [20,21]. Therefore, in most of the cases, quanti-
tative analysis represents the only method to differentiate various prov-
enances of these materials.
As a main result of this research, all the methodological develop-
ments have been applied in the interpretation of clay structures in care-
fully selected case studies on Italian and Bohemian Baroque paintings,
and on Early Renaissance preparation layers for gilding.
2. Materials and methods
2.1. Claybinder interactionsMaterials and experiments
2.1.1. Reference mineralogical samples
Reference expandable clay structures of smectites and different in-
terstratied structures of illite and smectite were obtained from Source
Clay Repository (Clay Minerals Society, USA) for the study of their
12 D. Hradil et al. / Microchemical Journal 125 (2016) 1020
interaction with organic binders in paints. All these pure reference
clays are white or grey, while in the painting technology, one can nd
clays of different colours. Therefore, we have added also the so-called
Armenian bolea very traditional type of red clay rich in smectites,
which has been used, for example, as a preparation layer for gilding; how-
ever, our smectite-rich red clay is from Sardinia, as the geographical loca-
tion of original historical site in Turkey has been forgotten [22]. The
colours of clay-based pigments are determined by the presence of iron,
which is either incorporated in the silicate structure of the clay mineral
(in green earths) or admixed in the form of iron hydro(oxides)hematite
and/or goethite (in various reds and ochres).
2.1.2. Experimental procedure
For the claybinder interaction experiments, reference samples con-
taining expandable structures listed in Table 1 have been employed. In
addition to natural Ca2+/Mg2+ form of SAz-1 smectite, the chemically
modied homoionic Na+ form was prepared for the comparison. In
the clay technology, Na+ forms of smectites are considered to be more
reactive; however, in nature, they are less frequent than Ca2+/Mg2+
forms. Besides the presence of various interlayer cations, there is also
a difference in the number of water molecules lling the interlayer
space, as it has been mentioned in the introduction. To prepare the
homoionic smectite (indicated as NaSAz-1), the natural smectite SAz-
1 was suspended in a 1M solution of sodium chloride (grade p.a., sup-
plied by Lachema Neratovice, Czech Republic). The equilibrium of the
cation between the solution and the solid phase was re-established by
changing the supernatant. The process was nished by dialysis using
distilled water; the homoionic sample was dried at 60 C and powdered
using agate mortar [23].
The reference clay samples were mixed with skin glue (GRAC spol.
s.r.o), linseed oil (Kremer Pigmente GmbH & Co. KG), and egg yolk
(commercial production), which belong to the most widely used
binders in the historical painting technology, representing proteina-
ceous binders (glue), oil binders, and emulsions of both components
(egg yolk). Firstly, the reference clay samples were homogenised in an
agate mortar. Then, the solution of the skin glue with distilled water
(0.81 g per 15 ml) was prepared and allowed to swell for about 4
hours. After a slight warming in a water bath (not exceeding 60 C to
avoid the conversion of collagen to glutin [24,25], it was mixed directly
with the clay. Oil and egg yolk were applied without any pre-treatment.
To achieve a good level of homogenisation of mixtures, following
Table 1
Reference mineralogical samples and their composition [23,39,40].
Name/Code Origin/Supplier
Kaolinite Source Clay Repository of the Clay Minerals Society, USA
KGa-1b (www.clays.org)
Illite-smectite Source Clays Repository of the Clay Minerals Society, USA
ISCz-1 (www.clays.org)
Illite-smectite Source Clays Repository of the Clay Minerals Society, USA
ISMt-1 (www.clays.org)
Smectite Source Clays Repository of the Clay Minerals Society, USA
SAz-1 (www.clays.org)
Na-smectite Homoionic sodium form of SAz-1
NaSAz-1
Red bole smectitic Kremer Pigmente GmbH & Co. KG, Germany
K40490
Red bole kaolinitic Self-collection, locality Hoenec near Chomutov, Czech Republic
H111
Red earth Kremer Pigmente GmbH & Co. KG, Germany
K40503
Hematite Bayferrox, Lanxess, Germany
Bay-140M
Quartz Calofrig Borovany, Czech Republic
Q
Mica-muscovite Garmica Mdnec, Czech Republic
MIC
Anatase SigmaAldrich
ATS
ratios were applied: 1:3 for clayglue mixtures, 1:1.5 for clayoil mix-
tures, and approx. 1:1.3 for clayegg yolk mixtures. Reecting the signif-
icantly higher sorption capacity of pure smectite, ratios of 1:10 or even
1:16 were applied in the case of SAz-1 or NaSAy-1 samples, respectively.
Finally, claybinder mixtures were applied each on two glass slides
using paintbrush and let to dry on air. Non-accelerated drying took
from several hours (in most of the cases) up to several weeks (mixtures
with oil). As a reference, pure suspensions of clays with distilled water
were prepared as well and applied on glass slides in a similar way.
After interaction and drying of the samples, one slide of each of the sam-
ples was subjected to dehydration experiment in the laboratory dryer
under the controlled temperature of 210 C for 1 hour. As it appears
from our previous experiments, naturally hydrated expandable clay
structures dehydrate easily already at temperatures up to 100 C [26];
when the temperature exceeds 200 C, the process becomes partially
or fully irreversible. It leads to the collapse of interlayer space, which
is accompanied by a characteristic shift of basal diffraction lines. The
aim of the dehydration experiments was to prove that organic
molecules have intercalated the interlayer space resulting in an
incomplete collapse. All the samples pure reference ones, the ones
interacted with binder and the afterwards dehydrated were measured
by pXRD.
2.1.3. Micro-samples from actual paintings
Two micro-samples from Italian 17th century canvas paintings at-
tributed to A. Zanchi and G. B. Langetti and two micro-samples from
the early 16th century paintings on wooden support were selected for
the purpose of differentiation of expandable clay structures in micro-
scale (Table 2, codes J1124, J1116, J0614, J1328). The layers of interest
were represented either by the red-brown clay-rich grounds (in the
case of canvas paintings) or the red clay poliment under the water gild-
ing in the case of Renaissance works of art on wood.
2.2. Materials for quantitative analysis
2.2.1. Reference mineralogical samples
To evaluate the limits of quantitative determination of individual
phases in clay mixtures via micro-pXRD, reference samples of kaolinite,
quartz, mica, hematite, and anatase were obtained from different sup-
pliers and model mixtures were prepared in various proportions (see
below). For comparison, reference red earths from historical mining
Mineralogical composition by pXRD (minor phases in parantheses)
Weakly disordered kaolinite (+ quartz, illite, anatase) [39,40]
Interstratied randomly ordered illite/smectite (70/30) (+quartz, feldspar,
kaolinite) [39]
Interstratied randomly ordered illite/smectite (60/40) (+ quartz, feldspar, calcite) [39]
Smectite group mineralCa-rich montmorillonite (+ quartz, mica, feldspar) [39]
Smectite group mineralNa-montmorillonite (+ quartz) [23]
Smectite group mineralmontmorillonite, kaolinite, anatase, hematite (+ quartz, illite,
goyazite)
Kaolinite, anatase, hematite (+ quartz, illite, goyazite)
Quartz, kaolinite, illite, hematite (+ anatase)
Hematite
Quartzfraction under 20 m
Muscovitefraction under 30 m
Anatase
 D. Hradil et al. / Microchemical Journal 125 (2016) 1020
Table 2
List of micro-samples of paintings selected for this study.
Code Artwork, place
J1124 St. John the Evangelist (canvas painting), National Heritage Institute, Regional Ofce in
esk Budjovice (Budweiss), Nm nad Oslavou Chateau, CZ
J1116 Tityos / Prometheus (canvas painting), Art Gallery in Ostrava, CZ
M0731 Portrait of a painter of miniatures (canvas painting), National Gallery in Prague, CZ
J0904 Crowning with Thorns IV (canvas painting), St. Nicholas Church, Lesser Town, Prague, CZ
J0614 Altarpiece of St. Martin (panel painting), Church of St. Martin - the Bishop of Tours, Lipany, SK
J1328 Mourning of Christ (polychromy on wood), Franciscan Convent in Brno, CZ
sites in Western Bohemia (Hoenec) and Bavaria (Troschenreuth) used
particularly for making grounds in Mid-European Baroque painting
were self-collected or supplied by Kremer Pigmente, GmbH & Co. KG,
Germany. Both these sites represent signicant historical sources of
red clays, commercially known as Czech bole and German (or Bavar-
ian) bole, respectively. It is highly probable that artistic workshops/
painters in the 17th century preferably purchase already primed
canvases, i.e. canvases already coated by the ground material (commer-
cially available), and therefore, only economically signicant sources
are reliable to consider within this comparative study [21]. Mineralogi-
cal compositions of reference samples are summarised in Table 1.
2.2.2. Preparation of powder model mixtures and model layers imitating
ground layers of paintings (mock-ups)
To evaluate the reliability of pXRD and micro-pXRD data for quanti-
tative phase analysis of preparatory layers, model pigment mixtures
were prepared, similar to those used in Mid-European Baroque paint-
ing. Model mixtures of clay-based red earths were created from com-
mercially available materials. Subsequently, they were weighed in the
desired proportions and micronized in the McCrone micronizing mill
with the addition of 10 ml of ethanol for 10 min. After drying, samples
for X-ray powder diffraction experiments were prepared by side-
loading method to minimise the effect of preferential orientation.
Experimental model layers were prepared from the powder model
mixtures by adding animal glue (0.6 ml of the glue solution to 1 g of
powder model mixture). After that, the suspension was mixed with
0.16 g of polymerised linseed oil (Umton, CZ) and a drop of honey ac-
cording to a historical recipe [27]. A thin layer of the prepared suspen-
sion was applied on a support (Si zero background sample holder or
wooden board). The whole procedure was designed to simulate the art-
work sample analysis as closely as possible. After drying, the sample
was mounted into a Huber goniometric head for micro-pXRD analysis.
2.2.3. Micro-samples from actual paintings
Two micro-samples from Baroque canvas paintings created by the
renowned Czech Baroque painter Karel krta were selected with the
aim to perform quantitative phase analysis of the ground layer and com-
pare the data with the results obtained on reference samples. Based on
preliminary investigation, it is known that both these paintings have
been created in the region of Central Europe and contain red-brown
earthy ground. The samples are listed in Table 2, codes M0731, J0904).
2.3. Analytical techniques
2.3.1. Conventional BraggBrentano X-ray powder diffraction
measurements
Diffraction patterns were collected with a PANalytical XPert PRO
diffractometer equipped with a conventional X-ray tube (CoK radia-
tion, 40 kV, 30 mA, line focus) and a multichannel detector X'Celerator
with an anti-scatter shield. X-ray patterns were measured in the range
of 4100 2 with a step of 0.0167 and 1050 s counting per step.
13
Artist Date of Layer of interest
creation
Antonio Zanchi Not known Red-brown ground (bottom layer)
(16311722)
Giovanni Battista Not known Red-brown ground (bottom layer)
Langetti
(16351676)
Karel krta 1635 Red-brown ground (bottom layer)
(16101674)
Karel krta 1673 Red-brown ground (bottom layer)
(16101674)
H.E.R. (Hans Khler) 1526 Red poliment under the gilding
Ulm workshop (?) Around 1500 Red poliment under the silvering
Conventional BraggBrentano geometry was used with the following
parameters: 0.02 rad Soller slit, 0.25 divergence slit, 0.5 anti-scatter
slit, and 15 mm mask in the incident beam, 5.0 mm anti-scatter slit,
0.02 rad Soller slit and Fe beta-lter in the diffracted beam. The duration
of the scan: ca. 13 hours.
2.3.2. Laboratory powder X-ray micro-diffraction
Micro-diffraction experiments were also collected using a PANalytical
XPert PRO diffractometer, which has been previously evaluated for appli-
cation in cultural heritage and forensic sciences [13]. In this study, a CoK
tube with point focus, an X-ray mono-capillary with diameter of 0.1 mm,
and a multichannel detector X'Celerator with an anti-scatter shield were
used. A sample holder was adapted by adding z-(vertical) axis adjustment
(Huber 1005 goniometric head). X-ray patterns were measured in the
range of 480 2 with a step of 0.0334 and 2200 s counting per
step. 2.5 mm anti-scatter slit, 0.04 rad Soller slit, and Fe beta-lter were
used in the diffracted beam. The duration of the scan: ca. 12 hours.
2.3.3. Quantitative phase analysisRietveld renement
Quantication of the experimental data was performed using the
Rietveld method [28]. As the clay minerals (such as, e.g., kaolinite) ex-
hibit a wide range of disorder (stacking faults in the layer structure),
the BGMN code was used for all calculations [29]. This programme in-
cludes a code which permits the use of structural models describing cor-
rectly the disorder models [30,31] based on a simulation procedure of
the software package DIFFaX [32]. The disorder model of kaolinite was
applied according to Ufer et al. [33] models of other reference materials
were described as standard Rietveld models [34]. Since the well-
ordered kaolinite was used in the experiments, the calculation of kaolin-
ite content was modied as proposed by Ufer [33] using the combina-
tion of the disordered model of kaolinite and the ordered standard
Rietveld one. Rened parameters comprised unit cell parameters for
all phases as well as size and micro-strain broadening parameters. Pre-
ferred orientation was rened only for mica (muscovite) and ordered
kaolinite. Spherical harmonics of the 2nd degree was used [35]. Addi-
tional rened non-structural parameters were sample displacement
error and a 6th-degree polynomial for background modelling.
2.3.4. Preliminary analysis of micro-samples from actual paintings
Each micro-sample, taken carefully from the painting during its res-
toration, was split into two fragments. One of them was analysed as it is,
the second was embedded in polyester resin and ground in cross-
section using the Kompakt 1031 grinding machine. Then, the observa-
tions of the layer stratigraphy were made in white reected light as
well as UV light (365 and 470 nm) using OLYMPUS BX-60 light micro-
scope equipped with a digital camera Olympus DP 70 and/or Zeiss
Axio Imager A.2 light microscope with a digital Olympus DP 75 camera.
Scanning electron microscopes (Philips XL-30 CP and/or JEOL JSM6510)
with detectors of back-scattered electrons (SEM), coupled with EDAX
(Philips) and/or INCA (JEOL) EDS detecting unit were used to describe
the elemental composition of individual layers and to indicate the
14 D. Hradil et al. / Microchemical Journal 125 (2016) 1020
presence of aluminosilicates. Measurements were carried out in a low
vacuum mode, which allowed analysis of samples without conductive
coating of their surface.
For the basic determination of the type of the binder, histochemical
tests were combined with infrared micro-spectroscopy. Fuchsin S and
methylene blue were used for identication of the presence of proteins
in the cross-sections, Lugol's Solution for the identication of polysac-
charides. Infrared spectra of cross-sections were measured in specular
reection mode using the infrared microscope Continuum coupled
with Nexus spectrometer (Thermo Scientic, USA) at the Polymer
Institute in Brno, Czech Republic (micro FT-IR). Spectra were recorded
in the region of 4000650 cm1 with the resolution of 4 or 8 cm 1
and analysed using Omnic 7.2 (Thermo Scientic, USA). If possible, pro-
teinaceous binders were further differentiated via MALDI-TOF/TOF
(Autoex Speed instruments, Bruker-Daltonics).
3. Results and disscusion
3.1. Correct interpretation of clay structures in paints
3.1.1. Basic distinction of clay minerals in microscale
It is well known that basic groups of clay structures (represented ei-
ther by one or by two tetrahedral sheets coupled with one octahedral
sheet within a layer, with interlayer space either empty or occupied
by cations (hydrated-exchangeable or xed)) can be differentiated
based on their characteristic basal diffraction lines. Their intensity can
be increased by preparation of oriented specimen by slow sedimenta-
tion of clay particles on a glass slide [2]. Results of our comparative
experiments showed that this fundamental characterisation can be per-
formed with micro-diffraction patterns as well, but with the following
limitations: (i) detection limits are inuenced by worse resolution and
lower signal/noise ratio, (ii) surface of the sample has to be at to
avoid shielding in low angles, (iii) artefacts that stem from the xation
of the micro-sample either in the embedding resin or by adhesive tape
directly to the holder may appear in the diffraction pattern, and (iv) rel-
ative intensity of the basal diffraction lines of layered silicates is signi-
cantly suppressed when measuring cross-sections instead of fragments,
as shown in Fig. 1. When fragments of paints (paint chips) are analysed
either from their top or bottom, i.e. the beam hits the sample perpendic-
ular to the paint layers, intensities of basal lines are increased as in the
case of an oriented specimen. In the case of cross-sections, the orienta-
tion of clay particles to the beam is just the opposite, i.e. they are
analysed from the side; consequently, the intensity of their basal
lines decreases. Therefore, it is advantageous to analyse the untreated
Fig. 1. The change of the intensity of kaolinite's basal diffraction lines (00l) with respect to the treatment of the sample measured as oriented specimen (1), untreated fragment (2), or its
cross-section embedded in polyester resin (3); Kkaolinite, Iillite, Hhematite, Aanatase, Ggoyazite.
fragments rather than their cross-sections, but only when the layer of
interest is accessible from the bottom or top of the paint chip.
3.1.2. Claybinder interactions in paints
To unambiguously identify expandable structures (e.g. smectites)
from the other ones, the standard procedure usually includes powder-
ing of clay material and its intercalation by specic organic molecules
(such as ethylene glycol) in order to generate highly characteristic
changes in the diffraction pattern of expandable phases [2]. However,
such procedure cannot be performed on painting micro-samples or
their cross-sections. In this study, we propose an alternative approach
for the differentiation of expandable structures in paints, which makes
use of an experimental description of claybinder interactions that
spontaneously took place in colour layers. For this purpose, it is neces-
sary to independently identify the type of organic material present in
the layer and describe its effects on the diffraction pattern of each of
the expandable structures. Hypothetically, this should lead to the recon-
struction of the original basal spacing before the interaction. The rst set
of experiments comprised the reactions of basic binder types (animal
glue, oil, egg yolk) with smectites (either pure or in association with
other minerals in natural earths) and with smectites interstratied
with illite (mixed crystals). This choice of binders encompasses protein-
aceous binders (animal glue), oils (linseed oil), and natural emulsions
containing both proteins and fatty acids (represented by egg yolk,
which contains, in contrast to the whole egg, a higher amount of fatty
components). When analysing actual painting, it is not always easy to
distinguish the original binder from organic materials, which could
eventually enter into the layer from the surroundings (e.g. from pro-
teinaceous insulation between the support and the ground or from the
secondary conservation treatments). Nevertheless, for the purpose of
interpretation of clay structures, it does not matter whether diffraction
shifts were caused by proteins from the original binder or proteins
which have contaminated the layer.
The most important nding brought by the experiments is that the
structure of smectite expands with proteinaceous binder (from approx.
1.5 nm to more than 1.6 nm), while the basal spacing slightly decreases
with oil and egg yolk (where oil component prevails) (Table 3, Fig. 2).
The effect of water molecules on the basal spacing value was deter-
mined by dehydration experiments under 210 C on samples with
glue and oil (egg yolk samples were excluded because they exhibited
similar behaviour to oil). As it can be clearly seen in Table 3, under
increased temperature, only the pure structures containing water
have collapsed to approx. 1.0 nm (in agreement with already published
data [26]), while the samples with proteins and oils only very slightly
dehydrated or their basal spacing remained xed (particularly in
 D. Hradil et al. / Microchemical Journal 125 (2016) 1020 15

Table 3
The results of claybinder interactions as indicated by changes in basal spacing (d).

d (001) values (nm) Ca/Mg smectite SAz-1 Red bole smectitic K40490 Illite/smectite (70/30) ISCz-1 Illite/smectite (60/40) ISMt-1 Na smectite NaSAz-1

Ambient conditions
Pure sample 1.54 1.56 1.19 1.22 1.27
With glue 1.72 1.67/1.53 1.19 1.29/1.20 3.13
With oil 1.50 1.49 1.14 1.20 1.49/1.28
With egg yolk 1.44 Amorph. (? 1.35) 1.13 1.18 1.32

After increased temperature treatment (210 C)

Pure sample 1.09 Amorph. 1.06 1.07 0.98
With glue 1.56 Amorph. (? 1.67) 1.09 1.10 2.97
With oil 1.46 1.42 1.14 1.17 1.28

samples with oil). However, at this moment, the mechanism of the pro-
cess remains unclear; the xation of basal spacing may be caused by in-
tercalation with organic molecules, but there can also be an inuence of
hydrophobic environment in oil media among the clay grains, which
prevents the dehydration to proceed.
The diffraction patterns of interstratied structures showed shifts of
diffraction lines which roughly correspond to the relative content of
smectite layers (40% or 30%, respectively), and consequently, they are
not easy to be recognised; 30% of smectite in the mixed crystal seems
to be the limit for unambiguous detection of the process of interaction
by pXRD. This is documented particularly by the sample ISCz-1 with an-
imal glueit is not possible to say whether the interaction proceeded
satisfactorily, as no measurable change of basal spacing took place and
the structure collapsed almost completely under increased tempera-
ture. The effect of intercalation starts to be more visible in case of the
IMt-1 sample with 40% of smectites.
The behaviour of homoionic Nasmectite signicantly differed from
other smectites; the interaction with proteinaceous binder caused the
exfoliation of its structure, which can be recognised by a very dramatic
shift of its interlayer space to 3.2 nm. Such values have never been
observed in any of the micro-samples from paintings, and therefore, it
is probably not a common case in natural earths.
The performed experiments indicate that it should be possible to dif-
ferentiate structures of smectites from interstratied structures in the
diffraction patterns of the micro-samples of paint layers. The differenti-
ation of smectites and chlorites (which exhibit no interaction with or-
ganic molecules) is possible in the presence of pure proteinaceous
binders (as, e.g., animal glue).
3.1.3. Application of the method to actual paintings
The results of the above-described experiments have been used for
the differentiation of clay structures in red-brown clay-based grounds
of Baroque paintings. As an example, a characteristic part of micro-
diffraction pattern obtained on a fragment of ground of a painting by
Antonio Zanchi (16311722) is shown in Fig. 3. Despite the worse reso-
lution of micro-pXRD, we have clearly distinguished two diffraction
maxima at 1.62 and 1.28 nm. For the 1.62 nm spacing, which cannot be
tted by diffraction pattern of any known mineral in the database, the ex-
planation seems to be clearit represents an organo-clay complex of pure
Ca/Mg smectite with proteins. The presence of proteinaceous component

Fig. 2. Signicant results of claybinder interactions in model samples: (A) change of basal spacing of reference Ca/Mg smectite (1) after the reaction with animal glue (2) and linseed oil
(3), and (B) exfoliation of the structure of homoionic Na smectite (1) after the interaction with animal glue (2).
16 D. Hradil et al. / Microchemical Journal 125 (2016) 1020
Fig. 3. Part of the diffraction pattern of the ground of the painting by Antonio Zanchi
(3) showing diffractions corresponding to reference organo-clay complexes of proteina-
ceous binder with red bole smectiticK40490 (1) and interstratied illitesmectite
with. approx. 40% of smectite (2).
Fig. 4. Red poliment under the silvering on the Early Renaissance polychrome sculpture (J1328) as can be observed on the damaged surface of the sculpture (left), on the detached fragment
(bottom right) and in the micro-section (top right); simplied description of the layer stratigraphy: +1 chalk ground, +2 red poliment, +3 silver foil, +4 organic glaze, +5 over-painting.
in the ground layer was additionally conrmed by micro-FTIR analysis.
The 1.28 nm spacing evokes the presence of Na smectite; however, it
would have to remain unaffected by the binder. That is, of course, not pos-
sible, because Na smectite reacts very easily in the presence of proteins
and exfoliates. It is probable that this spacing is correctly explained by
the presence of interstratied structure with a highly approximate ratio
of illite to smectite 6040%. These interpretations are based on compari-
son with the reference diffraction lines obtained on model mixtures,
which are depicted together with the diffraction lines of the studied
micro-sample in Fig. 3. The only remarkable difference is that the diffrac-
tion lines of the model mixtures are more diffuse, the two maxima are
convoluted. This is caused by the simultaneous presence of an already
interacted structure and the remaining unreacted structure. It indicates
that the process of interaction has not been fully nished within the ex-
periment duration.
The identication of an interstratied clay structure in Baroque clay-
based ground is exceptionally important. Based on our long-term exten-
sive study of grounds of paintings with different origin and period of
creation, the ndings of (possibly) interstratied structures are very
rare, thus representing a very good and exact indicator of a specic re-
gional provenance of the source material. We have already found the
diffraction maxima at the same position (approx. 1.3 nm) in several
other paintings of North-Italian Baroque authors (such as, e.g., J. B.
Langetti) [21]. Therefore, it may represent a highly characteristic feature
of North-Italian grounds applied in the 17th century, particularly those
originating from the school of Venice. In the earth pigments, interstrat-
ied clay structures have never been described before.
Another example is presented in Figs. 4 and 5. In this case, the method
was used for the distinguishing of smectites (expandable structures) and
chlorites (non-expandable structures) in the preparation layers for gild-
ing/silvering that are dated to the Early Renaissance periodspecically
on the 16th century silvering on polychrome wooden sculpture from
the Franciscan Convent in Brno, Czech Republic. Its diffraction line at
 D. Hradil et al. / Microchemical Journal 125 (2016) 1020
Fig. 5. Part of the diffraction pattern of red poliment under the silvering on the Early Renaissance polychrome sculpture (J1328) showing an overlap of 001 diffraction lines of expandable
smectite (S) and non-expendable chlorite (Ch), which is here interpreted as the presence of chlorite, since no expansion has occurred in the presence of proteinaceous binder; other
phases: Ggypsum, Gigibbsite, Ccalcite, Iillite, Aanatase, Hhematite, Agsilver, Clsilver chloride (corrosion of the silver).
d001 = 1.45 nm (Fig. 5) should be interpreted as representing chlorites
(e.g., clinochlore, typically with basal spacing at approx. 1.42 nm) rather
than smectites (typically at approx. 1.50 nm), because with collagenous
proteins (identied in the layer using MALDI-TOF/TOF), the smectite
structures should expand to more than 1.6 nm. The usage of a less valu-
able clay instead of the originally supposed smectite-rich Armenian bole
traditionally used for the gilding preparation layers [22] was further indi-
cated by the presence of clay micas, quartz, and especially by gibbsite (al-
uminium hydroxide). The presence of gibbsite was preliminary indicated
already by SEM-EDS, because of high Al/Si ratio, which is not typical for
any natural earth. Gibbsite can indicate either an admixture of organic
lake precipitated on gibbsite-based substrate, or red-brown bauxite
(raw material naturally rich in Al hydro-oxides) added with the aim to
improve the colour of the material. Completely, the same composition
of the preparation layer for gilding was found on the 16th century altar-
piece from Lipany, Slovakia (not shown). Both these works of art exhibit
stylistic similarities with the South-German artistic schools, therefore,
materials and technological similarities should be also expected. In con-
clusion, the composition of red earths used as preparation layers for
water gilding (the so-called poliments) seems to be more variable
than expected in the literature and inuenced by the availability of the
material.
3.2. Limits of quantitative phase analysis (QPA) of clay minerals by micro-
pXRD
3.2.1. Experiments on model mixtures and layers
When the clay-based pigments do not exhibit any regionally specic
characteristics recognisable by qualitative phase analysis (e.g., the
above-mentioned ndings of unusual interstratied structures in
grounds, or chlorites instead of smectites in water gilding poliments),
the differentiation of the origin of the material represents a complicated
issue. In our previous research [20,21], we have found out that red
Baroque grounds used in the region of Central Europe always contain
variable portions of layered silicates kaolinite and mica (illite and/or
muscovite), accompanied by quartz, hematite, and anatase. A wide
range of samples have been analysed and the results showed that the
relative contents of these phases differit is highly probable that
based on these criteria, the materials can be differentiated and associat-
ed with only two historical mining sites in Upper Franks, Bavaria (today
accessible at Troschenreuth), and Western Bohemia (today accessible at
Hoenec). Although the calculated differences seem to be signicant,
17
the exact values of accuracy of nonstandard measurements by micro-
pXRD are not known. To deal with this problematic issue, we have de-
cided to perform comparative measurements of model mixtures con-
taining the above-mentioned phases in different concentrations by
both conventional pXRD (in BraggBrentano reection arrangement)
and the micro-pXRD.
Table 4 summarises the weight fractions calculated by Rietveld
method and their comparison with initial weights. Estimated standard
deviations (esd) are given in parentheses for each calculated value.
They were determined by an implemented process in the BGMN soft-
ware [29]; therefore, they may vary according to the employed
software. For this reason, additional criteria were taken into account
in the process of evaluation of the QPA accuracy, as dened, e.g., by
Madsen et al. [36]. The rst one is called bias (Eq. 1), the second one
total bias (Eq. 2). Its minimum value is the criterion of accuracy evalua-
tion in the analysis of complex clay containing mixtures in the inter-
laboratory round robin tests known as the Reynolds Cup organised by
the Clay Minerals Society [37,38].
Bias wt:% Measured wt:%True wt:% 1
Total Bias abs Measured wt:%Truewt:% 2
Finally, the last criterion of accuracy evaluated in this study is the rel-
ative error (er). To evaluate the quality of the Rietveld t, an agreement
factor Rwp was used, dened as implemented in the BGMN programme
[29].
As it follows from Table 4, agreement factors Rwp as well as standard
deviations (esd) are systematically higher in the case of micro-pXRD in
comparison with conventional measurements. Bias and total bias values
follow the same trend, but in both cases, they are relatively low: total
bias of conventional measurements is 4.32 in average, while being
5.72 for micro-pXRD. In general, slight differences between weighed
and calculated values can be caused either by minor impurities of
anatase, quartz, and mica in reference natural kaolinite KGa-1b (see
Table 2), and/or by slightly lower homogeneity resulting from only
short (10 min) milling of coarse-grained quartz and muscovite pow-
ders. Both these imperfections were given by the strategy to imitate
the real (and therefore not perfect) mixtures in ground layers of paint-
ings. The lower homogeneity affects the micro-diffraction experiments
to a greater extent due to small irradiated volumes [13]. All of the
18 D. Hradil et al. / Microchemical Journal 125 (2016) 1020

Table 4
Weighed and calculated weight fractions of phases in model mixtures and layerscomparison of BraggBrentano geometry (BB) and micro-pXRD (-XRD).

Anatase Hematite Kaolinite Quartz Mica total

% (w/w) er (%) bias % (w/w) er (%) bias % (w/w) er (%) bias % (w/w) er (%) bias % (w/w) er (%) bias bias Rwp

Weighed 15.00 20.01 60.00 5.00 0.0

BB (powder) 17.38(10 15.9 2.4 20.56(11) 2.7 0.5 56.56(50) 5.7 3.4 4.95(6) 0.9 0.0 0.0 6.4 2.54
XRD (layer) 18.26(30) 21.7 3.3 20.46(33) 2.2 0.4 57.20(180) 4.7 2.8 4.14(24) 17.2 0.9 0.0 7.4 7.49
Weighed 10.00 20.01 59.99 5.00 5.01
BB (powder) 11.89(9) 18.9 1.9 20.92(11) 4.5 0.9 57.85(53) 3.6 2.1 4.97(7) 0.7 0.0 4.38(26) 12.6 0.6 5.6 2.62
BB (+binder) 11.76(9) 17.6 1.8 20.56(12) 2.7 0.5 57.07(59) 4.9 2.9 5.12(7) 2.4 0.1 5.49(31) 9.6 0.5 5.8 2.71
-XRD (layer) 11.72(23) 17.2 1.7 20.23(27) 1.1 0.2 59.50(170) 0.8 0.5 4.57(20) 8.6 0.4 4.02(44) 19.8 1.0 3.9 6.61
-XRD (powder) 13.02(26) 30.2 3.0 21.52(29) 7.5 1.5 55.70(160) 7.2 4.3 5.47(22) 9.4 0.5 4.30(48) 14.2 0.7 10.0 6.80
Weighed 4.99 10.00 65.01 10.00 10.00
BB (powder) 6.36(6) 27.4 1.4 10.38(8) 3.8 0.4 63.63(47) 2.1 1.4 9.94(9) 0.6 0.1 9.70(28) 3.0 0.3 3.5 3.46
-XRD (layer) 6.55(22) 31.3 1.6 10.57(24) 5.7 0.6 66.10(150) 1.7 1.1 11.30(23) 13.0 1.3 5.46(42) 45.4 4.5 9.1 7.63
Weighed 5.00 10.00 40.01 30.00 14.99
BB (powder) 6.06(6) 21.3 1.1 10.43(8) 4.3 0.4 38.69(45) 3.3 1.3 29.30(14) 2.3 0.7 15.51(25) 3.5 0.5 4.0 3.37
-XRD (layer) 6.05(20) 21.0 1.1 10.36(23) 3.6 0.4 40.40(110) 1.0 0.4 31.56(32) 5.2 1.6 11.66(39) 22.2 3.3 6.7 7.49
Weighed 4.99 10.00 45.01 20.00 20.00
BB (powder) 6.07(6) 21.6 1.1 10.38(8) 3.8 0.4 43.98(45) 2.3 1.0 19.70(11) 1.5 0.3 19.87(24) 0.6 0.1 2.9 3.58
-XRD (layer) 4.70(19) 5.8 0.3 9.63(22) 3.7 0.4 43.44(100) 3.5 1.6 21.30(30) 6.5 1.3 20.93(74) 4.7 0.9 4.5 7.47
Weighed 5.00 5.01 74.98 10.00 5.01
BB (powder) 6.45(6) 29.0 1.5 5.32(7) 6.2 0.3 73.95(47) 1.4 1.0 9.75(8) 2.5 0.3 4.53(22) 9.6 0.5 3.5 4.58
-XRD (layer) 6.23(22) 24.6 1.2 4.64(21) 7.4 0.4 74.77(96) 0.3 0.2 10.10(22) 1.0 0.1 4.27(39) 14.8 0.7 2.7 9.15

model samples were measured as pure powders by BraggBrentano
conguration and as model layers by micro-pXRD, i.e. the samples were
mixed with binder and applied by paintbrush (see Methodology). To
specify the potential effect of the binder, two more measurements
were performed on one of the model samples (the second one
in Table 4)BraggBrentano measurement of the sample mixed with
binder and ground again after drying, and micro-pXRD measurement of
the layer deposited on the support from aqueous suspension without
the binder. Resulting values show that regarding the BraggBrentano
geometry, the inuence of the binder seems to be negligible. Surprisingly,
in the micro-diffraction geometry, the addition of the binder led to the
decrease of total bias. Therefore, it is possible that the addition of binders
positively contributes to the homogenisation of the powder and, as a
consequence, suppression of the effect of heterogeneity.
3.2.2. Application to natural earths and actual paintings
Regarding the performed experiments which showed that the
values of total bias range between 3% and 6% for conventional Bragg
Brentano and 310% for micro-diffraction experiments, we have decid-
ed to compare the phase composition of reference earths from historical
mining sites at Western Bohemia and Bavaria (Table 5). Each of the
measurements, either BraggBrentano or micro-pXRD, was performed
twice, each time on a different portion of the material. Therefore, the
main differences are most probably caused by the heterogeneity of the
material (as indicated, e.g., by the varying content of quartz in H111
sample from Hoenec). It is also evident that while the major phases
contained in the reference materials were identied without any prob-
lems, the minor phases (such as, e.g., siderite or traces of illite and
quartz in the H111 sample) are not detectable by micro-pXRD. One of
the possible explanations (besides the already discussed heterogeneity)
may lie in the worse detection limit of the micro-pXRD, which is around
5 wt.% as determined in our previous studies [13]. Nevertheless, when
analysing major mineral phases of these reference earths, sufcient ac-
curacy is attained in comparison with the conventional BraggBrentano
geometry. It enables an unambiguous differentiation between the red
earths from these two historical sources.
As a next step, quantitative analyses of earth grounds on fragments
obtained from two paintings created by the renowned Czech Baroque
painter Karel krta (16101674) were performed (Fig. 6). The rst
one of them (M0731) is dated to 1635 (i.e., to the period, when, accord-
ing to historical evidence, K. krta shortly stayed in Saxony during his
travel from Italy back to Prague), and the second one (J0904) was
painted in Prague in the last years of the painter's life. It represents
one of the paintings of the Calvary cycle created for the St. Nicholas
church in Lesser Townthe most important High Baroque building in
Prague. As it can be clearly seen from results presented in Table 5, the
grounds of these two paintings differ and can be associated with the
German earth from Bavaria in the case of the painting created in Saxony,

Table 5
Differentiation of red earths from historical mining sites in Western Bohemia (today accessible at Hoenec near Chomutov) and Upper Franks (today accessible at Troschenreuth near Bay-
reuth): reference samples and micro-samples of Baroque paintings.

Sample code Anatase Hematite Kaolinite Quartz Illite Goyazite Siderite Rwp

% (w/w) % (w/w) % (w/w) % (w/w) % (w/w) % (w/w) % (w/w)

Reference red bole from Hoenec (Western Bohemia)

H111 (BB, powder) 7.53(9) 19.00(15) 59.85(56) 6.51(9) 1.97(21) 4.93(15) 0.23(6) 4.76
H111 (BB, powder) 9.03(8) 20.90(13) 61.54(46) 0.97(5) 2.60(13) 3.43(11) 1.54(54) 3.37
H111 (-XRD, layer) 10.06(17) 21.72(26) 66.23(86) 2.00(12) 4.46
H111 (-XRD, layer) 12.67(24) 19.62(37) 66.30(40) 1.40(24) 6.06

Reference red earth from Troschenreuth (Upper Franks, Bavaria)

K40503 (BB, powder) 1.01(10) 9.22(17) 42.29(59) 31.72(26) 15.75(27) 7.29
K40503 (BB, powder) 1.31(10) 8.61(21) 40.69(59) 31.51(31) 17.88(32) 7.86
K40503 (-XRD, layer) 1.32(10) 7.74(20) 42.03(64) 32.05(28) 16.87(30) 5.63
K40503 (-XRD, layer) 0.95(18) 9.58(43) 45.32(86) 33.81(54) 10.33(76) 7.55

Ground layers of paintings by Karel krta dated to 1635 (M0731-3) and 1673 (J0904-3)
M0731 (-XRD, fragment) 1.59(60) 8.10(220) 34.20(190) 32.80(150) 23.20(150) 10.54
J0904 (-XRD, fragment) 7.08(31) 10.29(55) 79.60(240) 1.77(29) 1.33(28) 11.02
 D. Hradil et al. / Microchemical Journal 125 (2016) 1020
Fig. 6. Raw materialred kaolinitic bole (H111) at the outcrop near Hoenec, Czech Republic (left), and its use as ground for the painting by Karel krta in 1673 (right, layer +1).
and with the Czech red bole from Hoenec in the case of the painting
created in Prague. This interpretation can be made despite the
facts that (i) Rwp values are signicantly higher when tting micro-
diffraction pattern of a real sample, (ii) total bias error is expected to
be up to 10%, and (iii) heterogeneity of mineral deposits results in
several differences in contents of, e.g., illite, kaolinite, or hematite,
when comparing Baroque materials with those available today. Further,
the above-described difference in mineralogical composition is in
agreement with differences in elemental composition obtained by
SEM-EDS. It can be demonstrated on characteristic elemental ratios,
as, e.g. K/Ti (=0.21 for J0904 and =11.52 for M0731 sample). There-
fore, we can conclude that the composition of the clay-based grounds
corresponds with the region of creation of the painting and/or with
the change of suppliers of the primed canvases by the artist in a partic-
ular place. They cannot be applied as a sign of authorship because within
the production of one painter, their compositions vary. The technology
of making grounds represents a much broader topic, where one should
consider also the mixing of materials from different sources or using of
additional colourants. As follows from our results, optimised methodol-
ogy based on quantitative micro-pXRD analyses could provide exact
discriminants for such comparative studies.
4. Conclusions
X-ray powder micro-diffraction (micro-pXRD) is the only microana-
lytical method that allows a reliable differentiation of individual clay
minerals' structures in colour and ground layers of paintings. Besides
the common limitations of micro-pXRD (such as, e.g., worse resolution,
small size of irradiated area, etc.), the following should be considered to
avoid misleading interpretation of the micro-diffraction patterns:
1. Expandable clay structures of smectites and different types of inter-
stratied structures with smectites are modied by interaction
with organic binders in the layer. The experiments showed that pro-
teinaceous binders cause an increase of their basal spacing (relative
to the content of smectite layers in the structure), while the presence
of fatty compounds (in oils or egg yolk) results in its very slight de-
crease. In both cases, a subsequent dehydration of the structure
was prevented.
2. The accuracy of quantitative microanalysis of clay minerals in mix-
tures with other minerals in earth pigments is characterised by
total bias values in the range of 310 wt.%, while in the case of con-
ventional BraggBrentano geometry, the total bias does not exceed
6 wt.%. However, a more critical issue seems to be the effect of het-
erogeneity of natural materials, which tends to be more pronounced
in small irradiated volumes.
19
As a specic sign of their origin, randomly ordered illitesmectite in-
terstratied structures were differentiated from pure smectites in paint-
ing grounds used by Baroque Venetian painters, and similarly, chlorites
were identied instead of previously expected smectites in preparation
layers for gilding/silvering in two Early Renaissance works of art,
belonging (regardless of their current location) to the South-German
origin. The accuracy of quantitative microanalysis was found to be suf-
cient for differentiation of red earths from two historical sources in Ba-
varia and Bohemia, respectively, both in reference samples and in
micro-samples of Baroque grounds of paintings by the famous Czech
painter Karel krta.
Acknowledgements
The authors would like to thank our colleagues in ALMA Laboratory
for their help in this research and also tpnka Hrdlikov-Kukov for
performing MALDI-TOF/TOF measurements. Thanks belong also to all
collaborating restorers (particularly Igor Foga and Mario Krl) for shar-
ing their expertise. The study has been supported by Czech Science
Foundation, project no. 14-22984S and by Institutional Budgets of Re-
search Institutions (RVO 61388980 and 60461446).
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