Characterization of organic media in the wall-paintings of the “Palace of Nestor” at Pylos,...

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Characterization of organic media in the wall-paintings of the “Palace of Nestor”at Pylos, Greece: evidence for a secco painting techniques in the Bronze Age

H. Brecoulaki a,*, A. Andreotti b,**, I. Bonaduce b, M.P. Colombini b, A. Lluveras b

a Institute of Greek and Roman Antiquity, The National Hellenic Research Foundation, Vas. Konstandinou 48, 11635 Athens, GreecebDipartimento di Chimica e Chimica Industriale, Università di Pisa, Via Risorgimento 35, 5626 Pisa, Italy

a r t i c l e i n f o

Article history:Received 14 June 2011Received in revised form13 March 2012Accepted 11 April 2012

Keywords:Palace of NestorGreeceLate Bronze AgeWall-paintingsa seccoEggPlant gumsAnimal gluePYeGC/MSGC/MS

a b s t r a c t

This paper presents the results of an investigation of organic binding media detected in samples from theMycenaean wall-paintings at the “Palace of Nestor” in Pylos (Western Messenia, Greece): samples datedfrom the Late Bronze Age (ca. 1200 BC). This is the first scientific study to be conducted on organicbinding media in Mycenanean painting, and it is a study that aims to contribute significantly towards ourunderstanding of the original painting techniques used during the Bronze Age. The analytical proceduresemployed for the detection and characterization of the organic substances preserved within the paintlayers (proteinaceous materials, lipids and plant gums), were based on chromatography/mass spec-trometry techniques (PYeGC/MS, GC/MS). The most significant results, highlighting an a secco paintingtechnique, are herein reported and discussed.

� 2012 Elsevier Ltd. All rights reserved.

1. Introduction

Thewall-paintings of the Mycenaean “Palace of Nestor” at Pylos,due to the large number of preserved fragments (approximately17,000), their high artistic quality and their unusual iconographicdiversity (Lang, 1969; Brecoulaki et al., 2008) (Fig. 1), representa unique pictorial corpus of the Late Bronze Age. The palatialcomplex discovered by Carl Blegen and his colleagues in 1939 islocated on the Englianos Ridge in Western Messenia and is one ofthe most important and best preserved Mycenaean palaces (Blegenand Rawson, 1966, 2001; Davis, 2010).

The first substantial cache of Linear B tablets ever to bediscovered on the Greek mainland, was uncovered by the excava-tors in the archives of the palace. The palace itself consisted ofa Main Building, a Southwestern Building, a Northeastern Buildingand a Wine Magazine (Fig. 2) e structures that stood until the sitewas destroyed by a fire in 1180 BC. Fragments of wall-paintings

were found both inside the palace e either in situ or fallen fromthe surrounding walls e and outside it, in dumps and field scatters(Lang, 1969). The present paper focuses on the chemical charac-terization, by PYeGCeMS, GCeMS (Andreotti et al., 2006; Lluveraset al., 2010), of the organic binding media sealed within the paintlayers of thirty-eight micro-samples which were taken from wall-painting fragments from various rooms at the “Palace of Nestor”.

Since antiquity, organic materials composed either of proteinsand/or plant gums have been employed extensively as bindingmedia to disperse pigments on polychrome artifacts and wall-paintings (Newman and Halpine, 2001; Colombini et al., 2004;Brecoulaki, 2006). The identification of organic materials withinpaint samples by means of chromatography/mass spectrometry isof great significance regarding the reconstruction of ancientpainting techniques: proteinaceous materials, lipids, resins, plantgums, and synthetic polymers can be identified in very smallsamples (<1 mg) (Colombini and Modugno, 2009). However, dueto the complexity of the matrix, the limited amount of analyzablematerial and the possible degradation of original materials, thisidentification remains a difficult task. In the present study, theabove challenges were overcome due to the large number andgood state of preservation of the analyzed samples, in conjunction

* Corresponding author. Tel.: þ30 2107273682.** Corresponding author. Tel.: þ39 0502219305, þ39 3385944223; fax: þ390502219260.

E-mail addresses: [email protected] (H. Brecoulaki), [email protected] (A. Andreotti).

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Journal of Archaeological Science

journal homepage: http: / /www.elsevier .com/locate/ jas

0305-4403/$ e see front matter � 2012 Elsevier Ltd. All rights reserved.doi:10.1016/j.jas.2012.04.018

Journal of Archaeological Science 39 (2012) 2866e2876


Fig. 1. Reconstruction of the Throne Room at the “Palace of Nestor”, by Piet de Jong (copyright: University of Cincinnati).

Fig. 2. Key plan of “Palace of Nestor” (drawn by John Travlos, adapted by William B. Dinsmoor, Jr.).

H. Brecoulaki et al. / Journal of Archaeological Science 39 (2012) 2866e2876 2867


with the development of sophisticated quantitative GC/MS samplepreparation and analysis procedures, allowing for a systematiccross check and verification of the obtained results.

Indeed, despite the large number of publications on thecomposition of plaster and pigments of Bronze Age wall-paintings,there is still a large gap on the subject of organic binding media,relating to a secco painting techniques (painting on a dry plaster,which requires a binding medium to attach the pigments onto thewall). In previous literature on Bronze Age murals, the a fresco(method of mural painting in which the artist must startapplying his colors on the wet plaster layer) is considered thedefinitive wall-painting technique, and there has also beena marked preference of frescoes to wall-paintings. But theassumption that Bronze Age murals are, by definition, frescoes isbased exclusively on wall-painting replication experiments andthe physical appearance of painting (Cameron, 1968; Evely, 1999;Chryssikopoulou et al., 2000; Brysbaert, 2008), as well as on theobservation of the stratigraphy of paint in cross sections and onthe calcium distribution within a paint layer (Perdikatsis et al.,2000; Dandrau and Dubernet, 2006; Brysbaert, 2007). However,as can be seen in recent technological studies (Dandrau, 2001: 62;Dandrau and Dubernet, 2006: 246; Brysbaert, 2007: 160), themicroscopical cross-sectional analysis of paint layers combinedwith elemental analysis (SEMeEDS/EDX), and the examination ofpigment penetration into the plaster as a criterion for identifyingbuon fresco technique has led to contradictory interpretations.Therefore, even though the possibility of a mixed secco/frescotechnique has been proposed, it remains a hypothesis rather thana confirmed analytical fact (Brysbaert, 2008; Brecoulaki, 2010).Prior to this paper, there has been no systematic analytical studyof the organic binding media in the Aegean Bronze Age. Further-more, there is still a limited understanding of the interconnec-tedness of paint layers and plaster. The aims of the present studyare as follows:

(a) To establish the first comprehensive analytical documentationfor the study of the painting techniques practiced during theLate Bronze Age, and to fill the gap in our knowledge regardingthe use of organic binding media in antiquity.

(b) To discuss the obtained data in relation to the analyticalmethods employed and to evaluate their possibilities andlimitations.

(c) To stress the art historical importance of a secco paintingtechniques in Aegean painting, in relation to the practice ofsimilar techniques in the Eastern Mediterranean during theBronze Age.

(d) To evaluate the data from an archaeological perspective, anddiscuss both the diversity of the identified binding media inrelation to their distribution in the various rooms of the palaceand their micro-chronologies.

2. Materials and methods

2.1. Archaeological paint samples

The majority of the selected samples were taken from variousrooms in the Main Building of the palace, Hall 64 in the South-western Building and the North-West Slope Dump (Table 1, Fig. 3).The sampling was performed at the Archaeological Museum ofChora (where the wall-paintings are stored), on solid coloredfragments, which did not join or belong to any of the preservedfigural compositions. The criteria for the fragments’ selection reliedmainly on their state of preservation (burnt or heavily damagedpieces were avoided), their physical color (a representative varietyof colors was ensured) and the texture of their surface (thick and

well preserved paint layers were preferred). Table 1 reports the listof the thirty-eight samples analyzed, the organic materials identi-fied and the type of analysis used for their characterization. All thesamples were analyzed with the GC/MS procedure for the simul-taneous analysis of the proteinaceous, lipid and resinous materials(Andreotti et al., 2006). A large number were also subjected to theprocedure that allows also the analysis of the saccharides fractionfrom the same micro-sample (Lluveras et al., 2010). When possible,different layers were mechanically separated using a scalpel andtungsten needles and analyzed separately in order to obtaininformation on the distribution of the saccharide materials(Bonaduce et al., 2007) in both the paint layer and the plasterlayer(s). Whenever the sample size was large enough, a smallfraction was also analyzed by PYeGC/MS (Bonaduce and Andreotti,2009) in order to identify polymeric material.

2.2. Methods

2.2.1. Chemicals and reagentsMonosaccharide standards: D-(þ)-galactose (galact), L-(�)-fucose

(fuc), L-(þ)-arabinose (arab), L-(�)-ramnose (ramn), L-(�)-mannose(mann), D-(þ)-xylose (xyl), D-(þ)-glucose (gluc), D-glucuronic acid(gluc ac.), D-galacturonic acid (gal ac.), all produced bySigmaeAldrich or Fluka. As internal standardmannitol (I.S)was usedin 99% purity (SigmaeAldrich).

Amino acid standards from SigmaeAldrich: alanine (ala), glicine(gly), valine (val), leucine (leu), isoleucine (ile), serine (ser), proline(pro), phenylalanine (phe), aspartic acid (asp), glutammic acid(glu), hydroxyproline (hyp). As internal standard for derivatisationnorleucine (nor) was used in 99% purity (SigmaeAldrich), and asinternal standard for injection hexadecane (ed) was used in 99%purity (SigmaeAldrich). Fatty acids standards, as well as theinternal standard for derivatisation, tridecanoic acid (C13) andhexadecane the internal standard for injection (ED) were suppliedby SigmaeAldrich (Milan, Italy).

2.2.2. InstrumentationGas Chromatography/Mass Spectrometry GC/MSwas performed

according to the procedure reported in the literature (Andreottiet al., 2006; Lluveras et al., 2010; Bonaduce et al., 2007). A 6890NGC system gas chromatograph coupled with a 5973 mass selectivedetector single quadrupole mass spectrometer equippedwith split-splitless and a 6890N GC system gas chromatograph (AgilentTechnologies, Palo Alto, CA) coupled with a 5975 mass selectivedetector (single quadrupole mass spectrometer) equipped withPTV injector, by Agilent Technologies, were used. The MS transferline temperature was 280 �C; the MS ion source temperature waskept at 230 �C; and the MS quadrupole temperature was at 150 �C.For the gas chromatographic separation, a HP-5MS fused silicacapillary column (5% diphenyl/95% dimethyl-polysiloxane,30 m � 0.25 mm i.d., 0.25 mm film thickness, Agilent Technolo-gies) with a deactivated silica pre-column (2 m � 0.32 mm i.d., J)was used. The carrier gas was used in the constant flow mode (He,purity 99.995%) at 1.2 ml/min. The mass spectrometer operated inthe EI positive mode (70 eV) and MS spectra were recorded both inTIC (total ion current) and SIM (single ion monitoring) mode.

The PTV injector and the chromatographic oven was pro-grammed as follows:

Lipids-Resins. The PTV injectorwasused in splitlessmodeat280 �Cand the oven was: 80 �C isothermal for 2 min, 10 �C/min up to200 �C, 200 �C isothermal for 3min,10 �C/min up to 280 �C, 280 �Cisothermal for 3min, 20 �C/minupto300 �C isothermal for 30min.Amino Acids. The PTV injector was used in splitless mode at220 �C, and the oven was: initial temperature 100 �C isothermal

H. Brecoulaki et al. / Journal of Archaeological Science 39 (2012) 2866e28762868


for 2 min, then 4 �C/min up to 280 �C, 280 �C isothermal for15 min.Saccharides. The PTV injector was used in splitless mode at250 �C and the oven was: 50 �C isothermal for 2 min, 5 �C/minup to 190 �C at, 190 �C isothermal for 20 min 5 �C/min up to280 �C, 280 �C isothermal for 15 min e for saccharides: 50 �Cisothermal for 2 min, 5 �C/min up to 190 �C at, 190 �C isothermalfor 20 min 5 �C/min up to 280 �C, 280 �C isothermal for 15 min.

For the Pyrolysis/Gas Chromatography/Mass Spectrometrya continuous mode microfurnace pyrolyzing injection system Pir-ojector (SGE, Austin Texas, USA) was used. The pressure in thefurnace was kept at 14 psi and the purge flow was 0.5 ml/min. Thepyrolysis chamber was connected through a PTV injector toa 6890N GC System Gas Chromatograph (Agilent Technologies, PaloAlto, CA, USA) coupled with a 5973Mass Selective Detector (AgilentTechnologies, Palo Alto, CA, USA) single quadrupole mass spec-trometer. The MS transfer line temperature was 280 �C; the MS ion

source temperature was kept at 230 �C and the MS quadrupoletemperature at 150 �C. The mass spectrometer operated in the EIpositive mode (70 eV) and the mass range was from 50 to 750 m/z.For the gas chromatographic separation an HP-5MS fused silicacapillary column (5% diphenyl-95% dimethyl-polysiloxane,30 m � 0.25 mm i.d., 0.25 mm film thickness, J&W Scientific,Agilent Technologies, Palo Alto, CA, USA) with a deactivated silicapre-column (2 m � 0.32 mm i.d., J&W Scientific Agilent Technol-ogies, Palo Alto, CA, USA) was used. The PTV injector was used insplit mode at 300 �C, the split ratio being dependent on the samplesize. The chromatographic conditions were: 31 �C isothermal for8 min, 10 �C/min up to 240 �C and isothermal for 3 min, 20 �C/minup to 300 �C and isothermal for 30 min. The carrier gas was used inconstant flow mode (He, purity 99.995%) at 1.0 ml/min. A few mgof samples together with 5 ml of hexamethyldisilazane wereinserted into quartz tubes (4 cm � 0.53 mm) and placed into themicrofurnace pyrolyzing injection system Pirojector operating at550 �C (Pyroprobe 500 Series by CDS Analytical Inc.).

Table 1List of the archaeological paint samples and their location, the identified organic materials and the type of analysis performed (GC/MS, PYeGC/MS).

Room/area ‘Palace of Nestor’ Sample Surface colour Organic materials identified Methods of analysisa

Protein Lipid Saccharide material GCeMS, PYeGC/MS

Room 1 P1.01 purple egg <LOQ e

004.05.01 blue egg <LOQ mixture PYeGC/MS006.02.01 red egg egg <LOD PYeGC/MS006.04.01 pl blue egg egg e PYeGC/MS006.04.01 prep e e e mixture PYeGC/MS007.15.01 brown e e <LOD PYeGC/MS

Room 3/4 Dr.17.03/04 yellow animal glue <LOQ <LOD PYeGC/MSRoom 4 P9.04 red e e mixtureRoom 5 0.21.75.05 pink e e not identified PYeGC/MS

0.26.13.05 yellow e e <LOQ PYeGC/MSRoom 6 P18.06 dark red egg egg fruit tree gum

P20.06 light pink egg egg mixtureP22.06 purple <LOQ egg <LODP23.06 pink egg <LOQ mixtureP103.06 red egg/animal glue egg e

P106.06 brown egg egg e

Room 17 P15.17 pink egg e mixture PYeGC/MSRoom 27/31 075.105.27/31 blue egg egg mixture PYeGC/MS

075.105.27/31 prep1 e e mixture075.105.27/31 prep2 e e mixture

Room 31 P63 blue egg egg not identified (PY) PYeGC/MS065.77.31 dark blue e e mixture PYeGC/MS069.56.31 blue egg egg e PYeGC/MS

Room 27/32 063.70.27/32 blue egg/animal glue <LOQ e PYeGC/MS063.73.27/32 blue egg/animal glue egg e

Room 43 080.09.43 light green e <LOQ mixture PYeGC/MS083.07.43 gray e <LOQ not identified PYeGC/MS

Room 46 P10.46 e e mixture PYeGC/MS105.65.46 pl brown e e not identified PYeGC/MS105.65.46 prep e e e <LOD105.66.46 brown egg <LOQ <LOQ PYeGC/MS112.93.46 orange e e <LOD PYeGC/MS

Room 64 P1.64 mauve <LOQ <LOQ <LODP7.64 dark red egg egg <LODP11.64 purple egg egg <LODP13.64 purple and blue e e <LODP118.64 purpleered e e <LOQ PYeGC/MS135.43.64 brown egg/animal glue <LOQ not identified PYeGC/MS144.22.64 blue <LOQ <LOQ not identified (PY) PYeGC/MS

North-West Slope Dump 179.122.NWS red e e <LOQ158.16.NWS blue e e <LOD PYeGC/MS179.268.NWS blue e e mixture PYeGC/MS

Outside the palace Dr.201.1959 pink and blue e e <LOQ PYeGC/MS

e ¼ not analyzed.<LOQ ¼ material content below the LOQ.<LOD ¼ material content below the LOD.mixture ¼ gum tragacanth þ fruit tree gum þ glucose containing material.not identified ¼ material content above the LOQ, but not identified by using the decisional scheme.pl ¼ paint layer; prep ¼ preparation layer.

a GCeMS was applied to all the samples and PYeGCeMS only to the samples reported.



3. Results and discussion

3.1. Analysis by GC/MS

3.1.1. Proteinaceous materialA total of 23 samples have been submitted to the analysis of

proteinaceous content (Table 1). Several analytical blanks havebeen performed allowing us to determine that an amount of 0.4 mgof proteinaceous material at a 95% of confidence level is the limit ofquantification (LOQ) and 0.3 mg is the limit of detection (LOD). Thus,only samples with a proteinaceous content higher than 0.4 mgcontain a significant amount of protein where the binder may beidentified. Table 2 summarizes the percentage contents of theamino acids for the 20 samples which contain a protein contenthigher than 0.40 mg. Fig. 4 reports the chromatogram acquired inSIM mode for the sample 075.105.27/31.

In general, the amount of the proteinaceousmaterial found in allthe samples is low. For the majority of the samples, the absence of

hydroxyproline indicates the absence of animal glue, except infive samples which contain this binder (Dr.17.03/04, P103.06;063.70.27/32; 063.73.27/32; 135.43.64). The amino acid percent-ages of the samples have been submitted to the principal compo-nent analysis and the score plots are reported in Fig. 5. Generally,the samples are located inside or close to the egg cluster, and thefollowing considerations can be drawn for the groups of samplescoming from specific rooms in the “Palace of Nestor”:

� Room 01: the amino acid profile of the samples P1.01,004.05.01 and 06.02.01 is coherent with that of egg;

� Room 03/04: the single sample Dr.17.03/04 contains mainlyanimal glue. It unequivocally presents a very high content ofglycine and the marker hydroxyproline;

� Room 06: the samples are close to the egg cluster, but highcontents of glycine cause a shift towards the animal gluecluster. Originally, the painting might have contained a smallamount of animal glue together with egg like in the sample

Fig. 3. Photomicrographs of the top surface of representative paint samples.

Table 2Percentage contents of amino acids and absolute amounts of protein. Al ¼ alanine, gly ¼ glycine, val ¼ valine, leu ¼ leucine, ile ¼ Isoleucine, ser ¼ serine, pro ¼ proline,phe ¼ phenylalanine, asp ¼ aspartic acid, glu ¼ glutammic acid, hyp ¼ hydroxyproline.

Sample ala gly val leu ile ser pro phe asp glu hyp

P1.01 10.4 13.6 8.1 9.0 6.4 8.4 2.5 4.3 17.6 19.9 0.0004.05.01 10.7 9.2 13.6 18.7 11.9 2.1 10.3 10.3 7.0 6.0 0.0006.02.01 9.6 10.8 13.3 14.9 8.3 2.6 9.9 5.8 16.0 8.8 0006.04.01 pl 12.0 12.8 10.0 10.8 6.8 5.6 14.7 4.6 16.0 6.8 0Dr.17.03/04 19.7 38.6 4.0 4.0 2.2 3.3 13.5 1.9 5.0 2.0 5.8P18.06 13.8 26.0 13.1 16.6 10.8 8.8 0.3 6.4 1.9 2.4 0.0P20.06 10.5 13.6 8.3 9.2 6.8 8.1 2.4 4.6 17.0 19.5 0.0P23.06 11.9 18.2 8.6 9.1 5.7 6.1 2.4 4.5 17.1 16.4 0.0P103.06 11.4 18.4 9.5 12.7 7.0 8.6 9.3 4.6 11.1 3.9 3.5P106.06 14.8 16.0 11.3 15.4 8.2 3.5 3.7 5.7 8.9 12.5 0P15.17 10.6 17.4 9.2 8.7 6.5 5.4 2.8 5.5 17.9 16.0 0075.105.27/31 13.7 27.8 11.6 14.1 8.7 5.0 10.2 2.6 2.1 4.2 0.0069.56.31 14.4 14.3 10.5 14.3 8.1 11.0 16.5 1.9 4.1 4.8 0.0063.70.27/32 8.2 18.2 6.2 10.8 4.9 15.8 5.2 4.2 13.6 12.4 0.5063.73.27/32 12.4 22.9 11.9 16.4 9.6 4.9 8.7 3.1 5.1 4.7 0.3105.66.46 11.1 11.0 6.4 14.8 7.5 8.1 4 6.9 9.2 21.0 0.0P7.64 15.7 22.4 14.9 16.2 11.0 5.8 0.5 7.3 4.1 2.4 0.0P11.64 18.9 18.4 16.6 18.9 14.4 1.7 0.3 8.7 0.4 1.9 0.0135.43.64 13.7 16.0 7.6 10.2 5.8 9.9 5.9 6.4 12.4 10.0 1.9P63 10.9 11.4 6.0 7.3 4.3 7.3 3.8 4.1 15.1 29.9 0.0



P103.06 (presence of hydroxyproline and high amount ofglycine). However, it cannot be ruled out that a biological attackmay have caused an increase of glycine;

� Room 17: sample P15.17 is very similar to the samples fromRoom 06;

� Rooms 27, 31 and 32: the samples (069.56.31, 063.70.27/32,063.73.27/32, 075.105.27/32) are close to the egg cluster andthe relatively high content of proline and/or of glycine causesa shift to negative value of PC1, thus towards the animal gluecluster. For the samples 063.70.27/32, 063.73.27/32, a mixtureof egg and animal glue is possible due to the presence ofhydroxyproline. The amino acid profile of sample P63 iscoherent with that of egg.

� Room 46: sample 105.66.46 contains egg;� Room 64: samples P7.64 and P11.64 present an amino acidprofile almost degraded but still comparable with that of anegg binder. Sample 135.43.64 contains also a small amount ofanimal glue.

The above analytical data indicate that the major organic binderdetected within the examined samples is egg, and only in a fewcases an admixture of egg and animal glue seems to have beenpracticed as well.

3.1.2. Lipid materialThe amount of lipid material is generally very low, often below

the quantification limit (2.5 mg): only 12 samples of the 24 samplesanalyzed show a significant amount of lipid material. All thechromatograms acquired are very similar one to each other andshow a major presence of palmitic acid and stearic acid. Fig. 6reports the chromatogram of sample 075.105.27/31. None of themolecular markers of plant resins and waxes are present.

The quantitation of fatty acids, for those samples with a signifi-cant amount of fatty acids, permits us to rule out the presence ofdrying oils. The profiles are generally compatible with the egg lipidand the average of the characteristic ratio values of the acids, A/P ¼ 0.2, P/S ¼ 1, SD ¼ 2.7, are consistent with the lipid of egg(Colombini et al., 2009; Andreotti et al., 2008).

3.1.3. Saccharide materialThe detection and quantitation limit of the procedure, calcu-

lated as the sum of the LOQ and LOD of the monosaccharides anduronic acids, are 0,56 and 0.26 mg, respectively. A total of thirty-four samples have been analyzed, and the results show that

eleven samples present a content of saccharide material at theblank level, while the glycoside content of all the other samples ishigher than the LOD (five samples) and often higher than LOQ(eighteen samples). The glycoside profile of these last twenty-twosamples is reported in Table 3. Quantitative relative percentcontent of sugars is reported only when the saccharide content ofthe sample is above the LOQ while the presence/absence of thesugars is indicated for the samples with a content between theLOD and LOQ. Fig. 7 shows the SIM chromatogram correspondingto the saccharide fraction of the sample 075.105.27/31, where themonosaccharides and glucuronic acids identified have beenevidenced.

The amount of polysaccharides in the samples is generally verylow, with a similar sugar profile in all rooms. The evaluation of theglycoside profile of the samples with a saccharide content higherthan the quantification limit has been performed using the deci-sional scheme proposed in the literature (Bonaduce et al., 2007)highlighting the following general conclusions:

� All of the samples contain seven sugars and two uronic acidswith the exception of samples 105.65.46, 021.75.5, P18.06,080.09.43 and 135.43.64. Although it is reported in the litera-ture that arabic, fruit tree and tragancanth gums have been themost used in antiquity in the Mediterranean Basin, the sugarprofiles of these samples do not match any of them. A possibleexplanation could be found in the use of a mixture of gums,such as tragacanth and fruit tree gums (containing fucose andmannose, respectively), but more research is needed to betterunderstand the degradation of polysaccharide binders, ofwhich not much is actually known, and to widen the availabledatabase of reference saccharide binders.

� The saccharide profile of sample P18.06 (fucose and galactur-onic acid are missing) can be ascribed to that of a fruit tree gumby using the decisional scheme (arabinose/yes; ramnose andglucuronic acid/yes; fucose and galacturonic acid/no;mannose/yes; xylose/yes / fruit tree).

� The absence of glucose in samples 021.75.05, 105.65.46, of gal-acturonic acid in sample 080.09.43, and of glucuronic acid insample135.43.64donot allow identifying thesaccharidematerialsource in any of those samples by using the decisional scheme.

� The sugar content of the preparation layers (samples 105.65.46prep, 006.04.01 prep, 075.105.27/31 prep1, 075.105.27/31 prep2)present a saccharide content above the LOQ, (except for sample105.65.46 prep that is a blank of the procedure) with the samesugar profile as the pertaining paint layer, but with a lessersugar weight percentage (w/w%). This datum suggests a pene-tration of the organic medium from the surface paint layer intothe plaster layer below, instead of an independent use ofglycoside material into the preparation of the plaster.

Considering that almost all of the samples contain glucose ina percentage above 20%, that is much more than the average valueof reference for tragacanth gum and/or fruit tree gum, it ispossible that another source of sugar e such as starch or honey e

may have been present as well. In the majority of the samples, thexylose relative amount is higher (samples 006.04.01 prep,065.77.31, 105.65.46 pl, 135.43.64, 075.105 prep1, 075.105.27/31prep1, 075.105.27/31 prep2, 179.268.NWS) or similar (samples004.05.01, 021.75.05, P18.06, P15.17, 076.105.27/31, P10.46) to thatof arabinose (xyl/arab � 1), while the reference plant gums,including fruit tree and tragacanth gum, present a xylose/arabi-nose ratio lower than 1. The evolution of the profiles of poly-saccharide materials during ageing has already been observed inthe literature without giving any explanation (Bleton et al., 1996;Bonaduce et al., 2007). The inversion of the ratio between xylose

e

e

e

e

e

e

e

ee

Fig. 4. Gas chromatogram acquired in SIM mode of the proteinaceous fraction ofsample 075.105.27/31: the profile is similar to the egg one, but with an high content ofglycine.



and arabinose could be due to unidentified degradationphenomena already reported in the literature (Bleton et al., 1996;Colombini et al., 2002; Bonaduce et al., 2007). Actually very littleis still known about the ageing of plant gum binders and theirdegradation processes (Bonaduce et al., 2007; Andreotti et al.,2008).

3.2. Analysis by PYeGC/MS

The pyrolysis of the samples reported in Table 1, confirmed thepresence of the proteinaceous, lipid and saccharide materials(Andreotti et al., 2009; Bonaduce and Colombini, 2003; Bonaduceand Andreotti, 2009), already identified by GC/MS analysis.

Fig. 5. Score plot (PC1 on the abscisse and PC2 on the ordinate) of the samples with a content of proteinaceous material higher than the LOQ, organized in groups according to theirlocation (room number) at the “Palace of Nestor”.



PYeGC/MS was also used to highlight the presence of polymericmaterials. Modern synthetic materials e clearly ascribable torestoration treatments e were evidenced in only two samples:samples 063.70.27/32 (Fig. 8a) and 004.05.01 (Fig. 8b). The presenceof methyl acrylate andmethylmethacrylate in sample 063.70.27/32lead to the identification of Paraloid B72, a material that has beenextensively used as awall-painting surface consolidant, by restorers.Another synthetic acrylic polymer, ormost likely amixture of acrylicpolymers, may be recognized in sample 004.05.01 based on thepresence of methyl metacrylate (Paraloid B44, or PlexiglumM345),butyl acrylate (Primal N560) and i-butyl metacrylate in the pyro-gram (Paraloid B67, or Elvacite 2045, or Plexiglum P24).

3.3. The use of a secco and tempera painting techniques at the“Palace of Nestor”

The analytical investigation of the samples from the “Palace ofNestor” has demonstrated that a secco and tempera painting

techniques were clearly practiced for the creation of Mycenaeanwall-paintings at Pylos. This new information revitalizes the long-lasting debate on the technology of prehistoric wall-paintings inthe Aegean, offering a reliable background for further under-standing and reconstructing the various stages of their manufac-ture. The presence of egg in ten samples as the solitary bindingmedium has confirmed the practice of egg tempera technique,while the extensive use of vegetable gums, including both gumtragacanth and fruit tree gum, shows that Bronze Age painterscombined a variety of organic materials with pigments beforeapplying them to the walls. A very interesting issue that hasemerged from the present research is the concurrent use ofmultiple bindingmedia during the painting process. In five samples(004.05.01, P18.06, P20.06, P23.06, 075.105.27/31) both egg anda mixture of plant gums (gum tragacanth and fruit tree gum) havebeen identified within the paint layer. It is not possible to deter-mine the exact role of each substance, but it seems likely that theywould have served different purposes, therefore suggesting thepractice of sophisticated painting techniques. Egg was probablyemployed as the major binding medium for the pigments, since itsbonding capacities are in general stronger than those of the gums.

Fig. 6. Gas chromatogram acquired in TIC mode of the lipid fraction of sample075.105.27/31.

Table 3Glycoside profile of the archaeological paint samples.

Sample Sugarsa Sugar content/mg

Xyl Arab Ramn Fuc Gal ac. Gluc ac. Gluc Mann Galact

004.05.01 22.7 21.1 1.5 0.7 3.6 1.6 30.7 5.4 12.9 1.70006.04.01 prep 22.5 9.8 2.2 0.9 1.6 4.0 44.7 7.1 7.1 0.95P9.04 14.2 30.4 4.1 4.7 0.2 6.5 23.4 10.3 6.2 5.2021.75.05 26.1 24.1 6.1 2.0 1.7 2.2 0.0 15.2 22.6 0.57026.13-5 e y y e e e e y y 0.35P18.06 22.7 21.6 7.8 0.0 0.0 7.2 16.7 5.6 18.5 3.8P20.06 13.4 18.6 9.3 10.2 5.8 6.3 16.7 7.6 12.2 4.9P23.06 14.4 20.3 8.7 6.8 1.8 2.1 23.1 3.4 19.5 5.7P15.17 17.0 16.2 2.5 2.0 0.4 2.7 39.0 1.6 18.6 6.8075.105.27/31 8.2 7.7 1.4 0.7 1.5 2.2 62.2 7.0 9.0 1.18075.105.27/31 prep1 16.7 10.7 1.9 0.6 1.0 1.2 53.3 7.6 7.6 5.08075.105.27/31 prep2 22.4 19.7 3.3 1.0 1.4 1.2 36.4 5.8 8.8 1.10065.77.31 25.1 5.0 0.7 0.4 2.7 5.8 44.6 4.1 11.5 1.30080.09.43 9.0 18.5 2.5 1.7 0.0 0.4 29.8 7.3 30.8 1.52083.07.43 pl 7.9 10.5 7.3 1.8 1.3 0.7 40.3 13.3 16.8 2.41P10.46 14.9 13.5 6.9 4.6 0.9 0.9 33.3 4.4 20.6 7.2105.65.46 pl 49.6 13.9 1.56 0.4 2.03 9.29 0.0 8.89 14.23 1.49105.66.46 y y y e y y e y y 0.36P118.64 e y y y e e e e y 0.38135.43.64 27.1 21.2 0.9 0.5 0.8 0.0 35.1 4.2 10.2 2.10179.122.NWS pl y y y e e e e y y 0.37179.268.NWS 23.5 14.4 1.4 1.7 1.6 3.2 38.6 7.3 8.3 1.64Dr.201.1959 e e e e e e y e e 0.44

a y/e indicates sugars with a quantity higher/lower than the detection limit in the samples with a content of saccharide material between the quantification and detectionlimit.

Fig. 7. Chromatogram (SIMmode) of the saccharide fraction of the sample 075.105.27/31.



Furthermore, we should consider that larger grained pigments suchas Egyptian blue that has been largely attested on the wall-paintings at Pylos (Brecoulaki, in press), would have requireda strong medium to bind it (Fig. 9).

The Pylian painters may have used eggs fromwild birds, such asgeese or ducks, although it seems likely that fowl (Gallus gallus)were already domesticated in Greece during the Bronze Age (West,1988). Plant gums could have been applied as a kind of superficialcoating on top of the paint layer to protect it, or even as an isolation

layer between the plaster and the paint layer or/and between thepaint layers themselves. The use of Acacia gum as an isolation layerhas been reported on Late Classical painting (Kakoulli, 2002). Wemay not exclude the possibility, however, of an intentional mixturebetween egg and plant gums in order to ensure more elasticity tothe paint layer. Gum tragacanth (an exudate of the Astragalus genus,such as Astragalus Gommiferus) consists of two fractions, trag-antahin which is water-soluble and bassorin which is insoluble inwater (Twiley, 1984; Mohammadifar et al., 2006). Unlike gumarabic which dissolves in water, gum tragacanth swells, forminga highly viscous suspension with colloidal properties. Certain wildspecies of the genus Astragalus are native to Greece, and Theo-phrastus mentions the existence of this plant in Achaia (HistoriaPlantarum, 9.1.3). The local provenance of this material must havebeen why Pylian painters preferred it to Acacia gum which theywould have had to import from Egypt. Recent analysis of the wall-paintings from Nebamun, dated to the 18th Dynasty (1292e1190)and therefore contemporary to the Mycenaean palace at Pylos,attested to an extensive use of vegetable gums, in particular Acaciaand Prunus species (Stacey, 2008).

In sample P18.06 fruit tree gum together with egg anda glucose based material have been identified. Although thesource(s) of glucose remain unknown, its constant presence in thegum-containing samples could be linked to a possible addition ofstarch or honey to the plant gums in order to eliminate brittlenessafter drying. A similar phenomenon has been attested onEgyptian painted artifacts from the Museum of Fine Arts inBoston (Newman and Halpine, 2001). In four samples (P103.06,063.70.27/32, 063.73.27/32, 135.43.64) both egg and animal glueare present. It is interesting to note that in those samples there isno evidence for plant gums. The presence of mixed media, such asanimal glue, egg, wax and plant gums have been previouslyreported in Egyptian art, both on wall-paintings (Palet and Porta,1990; Newman and Serpico, 2000; Newman and Halpine, 2001),and on wooden sarcophagus and cartonnage fragments (Chivariet al., 1995). In this last case, the presence of animal glue wasassociated with the preparation layers made of calcite, while eggand plant gum components were attributed to the paint binder(Scott et al., 2009).

The complex structure of the paint layer attested on numerousfragments from the “Palace of Nestor” conforming to the practice ofa secco techniques is emphasized by the frequent superimpositionof thick paint layers on top of colored grounds (Figs. 10 and 11),often observed in the paint cross sections (Figs. 12 and 13).

a

b

Fig. 8. Pyrogram of sample 063.70.27/32 obtained without derivatization with HMDS:1 methyl acrylate, 2 benzene, 3 methyl metacrylate (trace), 4 toluene, 5 ethyl meth-acrylate, 6 styrene. b: Pyrogram of sample 004.05.01: 1 methyl methacrylate, 2 butylacrylate, 3 i-butyl methacrylate.

Fig. 9. Top surface of coarse grinded Egyptian blue on top of an organic pink (wall-panting fragment from Room 1, “Palace of Nestor”). (For interpretation of the refer-ences to colour in this figure legend, the reader is referred to the web version of thisarticle.) Fig. 10. Flowers, detail in raking light on fragment from Room 53, “Palace of Nestor”.



Although organic binders of both animal and plant origin havebeen attested in the majority of the examined samples comingfrom inside the palace, in the fragments coming from the North-West Slope Dump no organic binder of animal origin has beendetected. Sample 179.268.NWS contained exclusively a mixture ofplant gums. Considering that those fragments belong to an earlierphase of the palace, before its final destruction, we may wonder ifworkshops of that period had prioritized the use of plant gumsrather that binders of animal origin. More samples should beinvestigated in the future from contexts outside the palace, inorder to confirm whether there was a change in the paintingtechnique during the Mycenaean era at Pylos. Furthermore, thefact that organic substances were not identified in all of thesamples from the palace may be attributed to the ageing andsubsequent deterioration of the organic binder, which obstructsits identification, or even to the possibility that both a secco anda lime-based technique were employed. Nonetheless, the secureidentification of organic binders in the majority of the samplesleaves no doubt that tempera and a secco painting techniqueswere widely, if not exclusively, practiced by Mycenaean painters

at Pylos, contrary to the general belief that the buon frescoconstituted the major painting technique of Bronze Age wall-paintings.

4. Conclusions

The analyzed samples from the “Palace of Nestor” at Pylos showa complex composition and the simultaneous presence of severalorganic materials, both ancient and modern. The binding mediawere successfully characterized by a combination of GC/MS andPYeGC/MS. Table 1 summarizes the obtained results for the totalityof the analyzed samples (thirty-eight) assembled by room/area ofthe palace. Solitary binding media was identified in eleven samplesand mixtures of multiple media, including egg, animal glue andplant gums in eighteen samples. Egg, sometimes mixed withanimal glue, and a mixture of vegetable gums (gum tragacanth þ-fruit tree gum þ glucose containing material) are the main organicbinders. These, although showing signs of ageing and degradation,are still present and positively recognizable, confirming that thepigments were dispersed and applied onto the walls using organicbinders. The detection of modern polymeric materials are ascribedto restoration treatments, aiming to consolidate the paint layers.

The use of egg tempera and a secco techniques is well attestedon fragments both from outside and inside the palace. In themajority of the rooms, it seems that multiple media were used inthe production of thewall-paintings, suggesting a commonpracticeof pictorial techniques for their creation, independent of their styleand iconography. The extensive analytical investigation of thepainting techniques in the Pylian wall-paintings has enhanced ourappreciation of the painted decoration of the palace and hassignificantly contributed to the study of the painting materials usedby Mycenaean painters during the second millennium BC. For thefirst time, tempera and a secco techniques have been securelyidentified for such an early period in the history of ancient Greekpainting. The importance of the results is further emphasized bythe fact that during the Classical and late Hellenistic periods thesame organic binders were still used almost exclusively by Greekpainters (Colombini et al., 2004; Brecoulaki, 2006; Brecoulaki et al.,2006), demonstrating the persistence of the pictorial techniquesand use of materials throughout the Hellenic world, from the LateBronze Age to the second century BC.

Nonetheless, there are still intriguing questions that cannot beadequately answered until many more binding media analyses areperformed:

Fig. 11. Female archer’s arm, detail in raking light on fragment from Room 31, “Palaceof Nestor”.

Fig. 12. Cross section of sample from Room 6, showing the superimposition of twopaint layers.

Fig. 13. Cross section of sample from Room 64, showing the superimposition of twopaint layers.



- What is the exact role of each organic substance identifiedwithin the paint layer(s) and the preparation/plaster layer(s)?

- Is the preference for a certain type of binding medium relatedto workshops, or specific periods?

- To what extent is the practice of a secco techniques at Pylosrepresentative of a common pictorial technique in the Aegean?

- Could the practice of a secco technique in the Aegean be relatedto a technological transfer from the East?

It is hoped that the present research will be applied to theanalysis of organic media not only inwall-paintings from the majorpalatial centers of the Greekmainland, but also to sites on Crete andThera where much larger numbers of wall-paintings have beenpreserved, in order to shed more light on this important aspect ofBronze Age painting in the Aegean.

Acknowledgments

The authors gratefully acknowledge the Wiener Laboratory ofthe American School of Classical Studies at Athens, the Institute forAegean Prehistory and the Department of Classics of the Universityof Cincinnati for funding the present research. Hariclia Brecoulakiwould like to express many thanks to Dr. Sharon Stocker and Prof.Jack Davis for their permission to study the material and theirsupport towards the completion of this study, and the 38thEphorate of Prehistoric and Classical Antiquities for their assistancein sampling at the Museum of Chora.

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