Microchemical Journal 112 (2014) 137–149

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Microchemical Journal
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Effect of photodegradation on the identification of natural varnishes by

FT-IR spectroscopy
C. Azémard, C. Vieillescazes, M. Ménager ⁎
Equipe Ingénierie de la Restauration des Patrimoines Naturel et Culturel – IMBE UMR 7263 CNRS – IRD 237 Université d'Avignon et des Pays de Vaucluse, 84000 Avignon, France

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

Article history: In this study, natural varnishes used in ancient painting techniques were recreated following ancient recipes.
Received 4 September 2013 They were irradiated by artificial sunlight and analysed by FT-IR all along the accelerated ageing. The main nat-
Accepted 21 September 2013 ural resins used were: sandarac, copal, colophony, mastic and dammar in alcohol, spirit turpentine and oil media.
Available online 1 October 2013
This study aims to understand and follow the evolution of characteristic absorption bands for the differentiation
of varnish types by FT-IR analyses. Kinetics and evolutions of the compositions of the varnishes during photo-
Keywords:
FT-IR spectroscopy
ageing were also studied. Photodegradation reactions lead to the loss of many characteristic absorption bands
Varnish and the apparition of new ones. To assess the specific fingerprints of different resins which do not depend on
Photodegradation the degradation state, results were analysed by statistical treatment (Principal Component Analyses). This
Terpenic resin multi-disciplinary approach leads to the identification of new criterions including the decrease of specificity
caused by the photochemical reactions.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction the nature of the artwork, it is crucial to identify the composition of

Natural varnishes are composed of natural resin dissolved in alcohol,
oil or turpentine spirit. These materials are used since mediaeval time
by artists to protect and embellish the painting [1], music instruments
[2] and pieces of furniture [3]. The recipes are numerous and various
resins were used through time [4,5]. Resins are mostly composed by ter-
pene molecules and can be classified in two groups depending on their
chemical compositions [6]. The resins chosen for this study are the prin-
cipal resins used by ancient paint masters in their varnish which spectra
show similarities in FT-IR. The “diterpenic resins” used were sandarac
(Tetraclinis articulata), copals (Araucariaceae or Leguminosae families),
colophony (distillated from Pinus resin) and different turpentine resins
(Pinaceae family) [7]. For the triterpenic ones, mastic (Pistacia lentiscus)
and dammar (Dipterocarpaceae family) were used and are still used by
painters and restorers. Shellac resin, glues, waxes or binders have very
specific spectrum bands and are not studied here. For this purpose,
Daher et al. recently publish an exhaustive comparison of infrared spec-
trum of all organic media used in ancient varnishes [8]. The degradation
of the resins by the action of light induces the yellowing and crackling of
the painting varnishes. The restorers usually remove the degraded layer,
sometimes only partially, and apply a new one. In order to choose the
appropriated solvent and not deteriorate the paint layer and respect
⁎ Corresponding author at: Equipe Ingénierie de la Restauration des Patrimoines
Naturel et Culturel – IMBE, 33 rue Louis Pasteur, 84000 Avignon, France.
E-mail address: matthieu.menager@univ-avignon.fr (M. Ménager).
the ancient varnish.
Various analytical techniques can help determine this composition.
ATR-far-infrared with principal component analysis has been used to
separate the resins in different classes and even identify the different
resins inside some of those classes [9]. Raman and FT-IR spectroscopies
also enable to have a good separation between the classes of resins
[8,10,11]. Studies by other techniques as High Performance Liquid
Chromatography (HPLC) [11–14], Gas Chromatography coupled to a
Mass Spectrometer (GC–MS) [15–18], or fluorescent spectrometry [10]
can also be carried out on paint varnishes to complete the identification.
Among all these analytical techniques, FT-IR is widely used in
conservation sciences for the characterization of organic materials.
This cheap, non-destructive and fast method can enable to determine
the type of material used (resin, wax, bitumen …) [19]. It can be com-
bined with complementary spectroscopic or chromatographic tech-
niques. Moreover, portable FT-IR instruments can be used in order
to fulfil on site measurement [20]. Various studies have been done on
fresh resins [8–12,19,21–26] and Table 1 gives the characteristic absorp-
tion bands previously described in literature.
However, only a few works deal with the study of degradation
process by FT-IR analysis [9,12,21–23]. Moreover, comparison of aged
resins to assess the dependence of identification criteria to degradation
processes has never been done. This question is crucial when analyses
deal with the identification of old samples from archaeological or artis-
tic media. Indeed, the identification of the different natural resins used
in varnishes is getting harder as the ageing takes place.
Varnishes can undertake different degradation processes: thermal
degradation, biodegradation and photodegradation. As noticed in

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138 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149
Table 1
Characteristic infrared absorption bands of different natural resins mainly used as varnishes from previous literature.
Resin Bibliography 3500–2650 cm−1
General [12,19,24,25]3050–2990 ν(\C_C\); 3070–3088 ν(=C\H);
2937 νxx(C\H), CH3, CH2; 2844 νx(C\H), CH3, CH2 1644 ν(C_C)
Colophony 2936 ν(C\H), 2870 ν(C\H)
Diterpenic
Sandarac [8,19] 2933 ν(C\H); 2873 ν(C\H); 2849 ν(C\H);
2646–2654 ν(C\H)
Copal 2933 ν(C\H); 2873 ν(C\H); 2849 ν(C\H);
2646–2654 ν(C\H)
General [12,19,21,24,25] 3446–3425 ν(O\H), 3079–2650 ν(=C\H)
augmentation with ageing, 3050–2990
ν(\C_C\),
2948 νxx(C\H), CH3, CH2; 2868 νx(C\H), CH3
Triterpenic
Mastic [8,19] 2949 ν(C\H), 2874 ν(C\H)
Dammar [26] 2950 ν(C\H)
Oil [25]
different works [21,27], photochemical reactions seem to be the
most important because paintings, instruments or pieces of furniture
are exposed to light daily. R.L. Feller points out that lighting in the

national gallery of London undergoes around 11 2 Mlxh (million lux
per hour), showing the potential importance of such reactions in the
varnish chemistry [28]. Terpenic resins generally photochemically
degrade by different kinds of oxidation reactions and polymerization
[11,22,23,25]. In this way, under artificial light, sandarac and copal
resins undergo a cross-linking process leading to high molecular weight
polymer [23]. Colophony is known to undergo aromatisation of unsatu-
rated cycle and many hydroxylations. Major components of mastic
and dammar triterpenic fraction undergo Norrish I reactions leading
to highly oxidized by products.
In this work, we will first discuss the identification of fresh spirit,
alcoholic and oil varnishes notably by using deconvolution of CH
and CO absorption bands. We will then look at kinetic aspects of the
photodegradation process. Afterward, we will discuss the identification
criteria of different aged varnishes. Finally we will propose the statistical
treatment of infrared spectrum data in order to have better identifica-
tion of the different resins.
2. Material and methods
The varnishes were fabricated following ancient recipes coming
from manuscripts and old books [4,5,29]. The chosen resins were pur-
chased with a special attention to the botanical certification. The study
was made on five resins, botanically certified, four diterpenic resins
(or derivate): sandarac (EMIGA, S.A.), Manila copal (EMIGA, S.A.), colo-
phony (Kremer Pigmente GmbH & Co KG), Venice turpentine (Kremer
Pigmente GmbH & Co KG) and two triterpenic ones: mastic (The Chios
gum mastic growers association) and dammar (EMIGA, S.A.). Linseed
oil was purchased from Lefranc & Bourgeois and the turpentine spirit
from ARTECH.
For alcoholic varnishes we made a solution at 150 g/L in ethanol. The
resin was grinded, and then added to the solvent, heated and stirred for
a few minutes. Oil varnishes were fabricated by adding the powdered
resin into nearly boiling linseed oil (Lefranc & Bourgeois) with a 1:2
weight ratio, stirring it until total dissolution (unless for copal and
sandarac whose solubility in oil is weak). Spirit varnishes were made
the same way than alcoholic varnishes with a 375 g/L concentration in
turpentine spirit [29].
Alcoholic, spirit and oil varnishes were done with all resins except
sandarac. In fact oil and spirit varnishes were not feasible for sandarac
due to the limited solubility of this resin in such solvents.
The FT-IR analyses were made with a Thermo-Nicolet AVATAR 360
FT-IR in transmission mode with OMNIC 32 software. All FT-IR spectra
1750–1600 cm−1 1500–580 cm−1 fingerprint region
1718–1690 ν(C_O), 1384 δ(\CH3),1448–1445 δxx(CH3); 1386 δx(CH3);
888 δ(CH2) out of plane (exomethylene func)
1697 ν(C_O); 1610–1615 1496, 1365,1275, 1239, 1151, 1130, 1107, 980, 910,
823, 707, 652
1694 ν(C_O); 1643 vinyl 1497, 1466, 1449, 1329, 1315, 1259, 1259–1263, 1236,
1213, 1153, 972, 909, 885–890, 856, 823, 789–792
1694 ν(C_O); 1643 vinyl 1466, 1449, 1329, 1315, 1259–1263, 1228, 1149, 889,
850, 795
1713–1706 ν(C_O); 1456–1455 δ(CH3), δ(CH2) 1378–1384 δx(CH3)
1658–1639 vinyl 1037–1035 ν(C\O\C) ether and ν(C\O) alcohol
1717 ν(C_O); 1650 vinyl 1458–1460, 1245, 1161, 1115, 1046, 1008, 837, 580
1713ν(C_O); 1647 vinyl 1459, 1379 δx(CH3), 1307 ν (C\H), 1251 ν (C_C),
1180 and 1154 ν (C\C)sat or (C\H)aromatic ring,
1083 and 1032 ν (C\O)
1750–1730 ν(C_O) 1418 ν(C\O)
were collected in the middle infrared (400 to 4000 cm−1) recording
64 scans per spectrum with a data spacing of 1.929 cm−1. The samples
were prepared by coating KBr pellets with 10 μL of the varnish and then
dried at ambient temperature before analysis. All spectra presented or
used in this paper were baseline corrected and smoothed. Besides we
removed the CO2 band due to atmospheric conditions. Deconvolutions
were done with OMNIC 8.1 software, choosing a Gaussian pointing
and a medium sensibility.
The samples were irradiated in a Suntest CPS+ photoreactor (Atlas)
equipped with a xenon lamp and a filter that prevents the transmission
of wavelength below 290 nm. The lamp was set at the intensity of
750 W·m−2. The temperature of the sample was roughly maintained
at 35 °C. The pellets were periodically analysed by FT-IR to follow the
photodegradation process. Similar experiments were led on glass plates
and gave the same results showing the absence of reactivity of the KBr
pellet.
The Principal Component Analyses were performed on the whole
spectrum with TQ analysis software using mean centering data with a
covariance algorithm.
GC–MS analyses were performed with a Thermo Fischer device
equipped with a GC Focus coupled with an ITQ700 mass spectrometer.
Samples were previously thimethylsilylated by using BSTFA (Aldrich);
3. Results and discussion
Each spectrum can be divided in four zones where information for
identification can be found. The different regions are the OH region
(3700–3200 cm−1), the CH region (3200–2400 cm−1), the carbonyl
region (1800–1550 cm−1) and to finish the fingerprint region (1550–
500 cm−1).
Samples made with a resin dissolved in spirit turpentine or ethanol
show the same infrared spectra and degradation kinetics (Appendix
A). Moreover, Venice turpentine varnishes lead to similar infrared spec-
trum and degradation process than colophony. Thus, these results do
not appear in our paper but are available in the appendices. Infrared
spectrum of fresh and 800 h irradiated varnishes are given in Fig. 1(a)
and (b).
3.1. Identification of fresh varnishes
3.1.1. Alcoholic and spirit varnishes
3.1.1.1. CH region. In non-aged diterpenic alcoholic varnishes, the
3080 cm−1 band corresponding to a C_C double bonds stretching
was previously described (Table 1) as a characteristic band for the
vinyl function of pimarane in Coniferae resins [8]. This band was used
 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149 139

Fig. 1. Infrared spectrum of alcoholic varnishes: fresh (a) and irradiated during 800 h in Suntest (b).

as a criterion of identification for diterpenic resins but it is also present For all fresh samples, spectra have similar CH region with maximum
in our dammar varnish showing that it can appear in triterpenic resins at 2933–2935 cm−1 for diterpenic varnishes and 2943–2948 cm−1
too. for triterpenic varnishes. As clearly shown on Fig. 2 for dammar and

Fig. 2. Deconvolution of the CH band of colophony and dammar (3200–2810 cm−1).

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C. Azémard et al. / Microchemical Journal 112 (2014) 137–149
Fig. 3. Deconvolution of the carbonyl band for the different resin studied.
 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149
Fig. 4. FT-IR spectra of linseed oil and oil varnishes: fresh (a) and irradiated during 800 h in Suntest (b).
colophony, a deconvolution of the signal between 2850 and 3050 cm−1
shows five absorption bands for both diterpenic and triterpenic resins
due to CH3, CH2 and tertiary CH stretching. It is worth noticing that a
sixth band centred on 2836 cm−1 (CH2 stretching) is only present in
diterpenic resin, and could be attributed to the presence of pimarane's
vinyl functions. However, this band is too weak to be a good identifica-
tion criterion. Moreover, CH3 stretching absorption band moves from
2955 cm−1 in diterpenic resin to more intense one centred at
2963 cm−1 in triterpenic resins. This gives the particular shape and ab-
sorption maximum to mastic and dammar resin spectra. This effect is
well linked to a triterpenic varnish chemical composition that shows a
higher proportion of methyl groups by molecule (around 25 wt.% in
lupenes, oleanenes, ursenes, dammarenes, tirucalenes [30]) than
diterpenic one (around 15 wt.% in abietanes, pimaranes, labdanes [31]).
Fresh colophony and Venetian turpentine alcoholic varnishes have a
very particular shape. Infrared spectra show a broad weak double struc-
tured band with maximum at 2652 and 2536 cm−1. This signal is
Fig. 5. Evolution of colophony infrared spectrum during the irradiation.
141
characteristic of absorption of hydrogen bonded OH group from carbox-
ylic function [32].
3.1.1.2. Carbonyl region. The carbonyl region also presents a characteristic
fingerprint, the band appears around 1703 cm−1 for triterpenic resins
and 1693 cm−1 for diterpenic resins. It is interesting to apply a
deconvolution on those bands to have a better understanding of this dif-
ference. Fig. 3 shows the presence of 3 main absorption bands between
1760 and 1820 cm−1. For diterpenic resins, a higher absorption band
with a maximum around 1720 cm−1 can be attributed to C_O bonds
in carboxylic group linked to saturated carbon ring in good agreement
with literature [32]. Indeed, chemical composition includes many carbox-
ylic groups linked to the carbon 4 of main diterpene molecules (abietane,
labdane, pimarane) with almost no ketonic carbonyl group in struc-
tures [31]. For triterpenic resins, the first absorption band is shifted at
1730 cm−1. It could be assigned to the saturated ring ketone as principal
triterpene biomarkers (for example dammarenone, amyrone, oleanonic,
moronic, masticadienoic acid) include such functional group on carbon
3. Absorption bands around 1640 cm−1, attributed to linear C_C
stretching vibration, are present in four resins: dammar, mastic, Manilla
copal and sandarac. This could be explained by the presence, in these
resins of unsatured polymer. Indeed, Manilla copal and sandarac contain
communic acid-based polymer [23], mastic resin is partly composed of
cis-1,4-poly-β-myrcene [33] and dammar resin of polycadiene [30].
3.1.1.3. Fingerprint region. The fingerprint zone shows many differences
between different samples. The diterpenic resins present a weak band
at 1410 cm−1, which also appears in oil varnishes spectrum, and
have small bands around 1445 cm−1, 1235 cm−1, 975 cm−1 and
793 cm−1 that were observed in previous literature (see Table 1). It is
important to point out that absorption bands centred at 1201 cm−1
and 805 cm−1 are characteristic of diterpenic resins. Colophony and
rosin also present specific bands at 1276 (C\O\ [32]) and 709 cm−1
(cf. Table 1). It is likely that all resins coming from Pinaceae family are
chemically close and cannot be differentiated by IRTF. Manila copal
has a specific band at 1035 cm−1 (C\O\C [24]) and sandarac at 1321
and 673 cm−1. The spectra of the two varnishes based on triterpenic
resin are very similar. However, few bands differ in the fingerprint
region: dammar possesses bands at 1303 (C\H) and 1286 cm−1
(C\O) and mastic a band at 1158 cm−1.
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C. Azémard et al. / Microchemical Journal 112 (2014) 137–149
Fig. 6. Kinetic evolution of different bands (a. C\O band, b. C_C band for copal and sandarac, c. C\H band) for the resins relatively to the absorbance of the fresh varnish (1: sandarac, 2: copal, 3: colophony, 4: mastic, 5: dammar varnishes; 6:
colophony oil varnish).
 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149 143

Table 2
Characteristic bands of the different fresh and aged resins and oils used in the varnishes (specific of Dit: diterpenic resin, Trit: triterpenic resin, Sand: sandarac, Col: colophony, Vt: Venice
turpentine, Dam: dammar, Mast: mastic, Ag: Ageing or degradation marker).

Diterpenic resins Triterpenic resins Linseed oil

Sandarac Copal Colophony/Vt Mastic Dammar

Wavenumber
Fresh Aged Fresh Aged Fresh Aged Fresh Aged Fresh Aged Fresh Aged
(cm–1)

3080 x x X x

3008 Oil

2963 Dit Dit Dit Dit Dit Dit

2940–2950 Trit Trit Trit Trit

2930–2940 Dit Dit Dit Dit Dit Dit

2855 Oil Oil

2836 Dit Dit

2530–2650 Col/Vt Col/Vt

1740 Oil Oil

1730 x x Trit x Trit x

1718 Dit Dit Dit

1445 Dit Dit Dit

1410 Dit Dit Dit Dit Dit

1321 Sand

1303 Dam

1286 Dam

1276 Col/Vt Col/Vt

1235 Dit x Dit x Dit x x x

1201 Trit Trit

1178 Ag Ag Ag Ag Ag Ag

1166 Oil

1158 Mast

1102 Oil Oil

1035 x Copal x x x x

975 Dit Dit Dit Dit Dit

805 Trit Trit

793 Dit Dit Dit Dit

723 Oil Oil

709 Col/Vt Col/Vt

673 Sand Sand

3.1.2. Oil varnishes
Oil varnishes can be easily identified thanks to the absorption bands
characteristic of oils. Oils have strong absorption band at 3008 cm−1
corresponding to C_C double bond stretching, 2856 cm−1 due to
ν(C_H) vibration, 1170 cm−1 which is strong and broad, 1102 cm−1
and a really specific one at 723 cm−1. Besides the carbonyl band
appears at a higher wave number than for the alcohol varnishes.
Because of the strong influence of the oil it is very difficult to identify
the resin employed in these varnishes. Among the three oil varnishes
that were studied only the colophony based one can be distinguished
as it presents some of the colophony characteristic bands around 2530
and 2650 cm−1 (Fig. 4a).
144 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149
3.2. Kinetic study
Spirit and alcoholic varnishes undergo the same degradation kinet-
ics. The evolution during ageing can be divided in three phases:
- A drying process that goes on for 5 to 15 h (24 h for oil varnishes)
and is marked by the strong change of the spectrum (cf. appendix C).
- A fast ageing process which goes on for 300 h (phase I).
- A long ageing process between 300 and 800 h where there is only
straight variation of the infrared spectrum (phase II).
First of all, it is very important to point out the similarities in the
photodegradation kinetics for all the varnishes. This clearly shows that
the photochemical processes involved in the different degradations
are similar for all the different natural resins used. We will then discuss
the evolution of infrared absorbance general to all varnish layers. The
example of the evolution of colophony infrared spectrum during the
irradiation is given in Fig. 5.
First, there is an augmentation of the absorbance between 3200 and
3600 cm−1 caused by the augmentation of OH bonds certainly due to
oxidation reactions. Furthermore, the absorbance of the fingerprint
zone increases between 1330 cm−1 and 800 cm−1 due to C\O
stretching absorption. As shown on Fig. 6a, at 1180 cm−1, we can see
that absorbance increases during approximately the first 300 h and
then remains almost the same or weakly decreases for all resins. At
the same time the absorbance of C_C band at 1644 cm−1 drastically
decreases especially for sandarac and copal (Fig. 6b). Finally, the carbon-
yl band broadens for all resins during artificial ageing. All those kinetics
show the importance of oxidation reactions, leading to the hydratation
of di and triterpene double bonds and hydroxylation of triterpene keto
group.
Fig. 6c clearly shows the decrease of the C\H band at 1455 cm−1,
this can be also been observed at 2935 cm−1 and 889 cm−1. This effect
is probably caused by the loss of volatile compounds (like sesquiter-
penes), low molecular weight terpenic degradation products and degra-
dation products of the polymeric fraction as already observed in other
synthetic polymer [34]. Moreover, global diminution of C_C and C\H
bonds can be also explained by the cross-liking, mostly in copal and san-
darac with formation of polycommunic acid as already mentioned by
Scalarone et al. [23].
Some other evolutions are more specific: in the case of colophony, the
1380 cm−1 absorption band (C\O) increases during drying process like
the other resins but then it decreases with the second process (5–800 h).
Another particular evolution is the one of the carbonyl band. Indeed its
absorbance slightly or strongly decreases or stagnates for diterpenic
resin varnishes whereas it increases for triterpenic ones.
Finally, we also can conclude that 800 h irradiated varnishes are
suitable to study aged varnishes. Indeed, there is a continuous evolution
of infrared spectrum between 0 and 800 h. Thus, if we want to find per-
tinent infrared fingerprint of resin, we have to search for absorption
band present all along the irradiation.
3.3. Identification of aged varnishes
3.3.1. The alcoholic and spirit varnishes
Evolution of the characteristic bands during irradiation is reported in
Table 2. It is important to note that many characteristic bands disappear
during the ageing process. Yet after artificial sunlight irradiation, the
stretching band around 2940 cm−1 remains visually different between
diterpenic and tritepenic resins. In fact, diterpenic resins show a shoul-
der at 2953 cm−1 and triterpenic ones have a maximum around
2950 cm−1 similar to fresh resins. On contrary the carbonyl band
loses its specificity as the band gets broader. The complexity of the
C_O absorption band combination makes any interpretation very diffi-
cult and even deconvolution is not appropriate to distinguish the differ-
ent types of resins.
In all alcoholic and spirit varnishes, the fingerprint zone shows
the same evolution: the bands at 1445 cm−1 (\CH bands [11]),
1201 cm−1 (δ(C\H) and δ(OH) [11]) and 805 cm−1 (C_CH
band [11]) disappeared. All these varnishes present a band around
1235 cm−1 (CO band) with a shape specific to the type of varnish. The
1410 cm−1 (COOH [32]) and 975 cm−1 bands are still present for
some of the diterpenic varnishes and the 793 cm−1 for sandarac.
Colophony varnishes keep some of their characteristic during ageing
(double band at 2534–2649 cm−1, bands at 1275 and 709 cm−1).
They can also be identified thanks to their C\O bonds absorption
band which gets broader forming a non-Gaussian band unlike the
other resins. For sandarac varnishes, only the characteristic 673 cm−1
(C\C band [11]) band remains after 800 h of irradiation. Moreover,
a good fingerprint of the degradation state of all types of varnishes
is given by the band around 1180 cm−1 (C\O bonds). Its presence
and intensity is well linked to the oxidation level of resin or the oil.
To conclude, identification of chemical composition of an unknown
sample by the research of specific absorption band is only possible by
(i) identification of the resin type (diterpenic or triterpenic) with the
specificity of the CH absorption and (ii) combination of absorption
bands specific and non specific band to a resin family according to
Table 2. For triterpenic resins, no precise identification is possible after
a certain degradation stage.
Varnishes sampled from painting could have various levels of degra-
dation. So, most of the time, analysis of these samples gives results
mixing molecular fingerprint of fresh and aged resins. Identifications
can, thus, be based on both fresh and aged characteristic bands given
in Table 2.
3.3.2. The oil varnishes
The oil varnishes spectra keep their specific shape through ageing
(Fig. 4b). Though the 3008 cm−1 band disappeared during the irradia-
tion, 2856 cm−1, 1102 cm−1 and 723 cm−1 bands remain with some-
times a small shift. The resin signal is weaker than for fresh varnishes
and it is more complicated to identify the resin employed. The initial
ratio in oil (2:1) was found in numerous recipes [5,35,36] and different
preliminary lab experiments show that it is very difficult to dilute a
higher mass of resin in oil.
Fig. 7. 3D visualization of all alcoholic varnishes by PCA with principal component1, 2
and 4 on the axes (sandarac: blue, copal: green, colophony: orange, mastic: red, dammar:
black).
 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149 145

Fig. 8. Principal component analysis of a) all our varnishes, b) alcoholic varnishes.

3.3.3. Principal component analysis of infrared spectrum
In order to improve the FT-IR identification of aged varnishes, we
applied statistical treatment by Principal Component Analysis (PCA).
All spectra were considered between t5 h and tmax to eliminate the
variability link to the drying process. The study of the FT-IR results by
Principal Component Analysis (PCA) shows a separation of the different
varnishes. We chose the most significant principal component scores,
PC1 and PC2 (and PC4 for 3D representation), for the statistical analysis
of our samples. Each point represents an FT-IR analysis of a varnish at a
particular time of irradiation between 5 and 800 h.
At first, the PCA was built from the whole spectrum from 400 to
4000 cm−1 of alcoholic varnishes. As shown on Fig. 7, on the 3D

Fig. 9. Principal component spectra of PC1 and PC2 of the alcoholic varnishes analyse.
146 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149
representation (PC1, PC2 and PC4), it is noticeable that the spectra
become less and less separated during the irradiation. The degradation
provokes a standardization of the spectra which is obvious in such
representation. The most discriminated varnish is the colophony one,
probably because of the specific bands previously mentioned.
As shown on Fig. 8a, the oil varnishes are well separated from the
alcoholic ones by this method. Moreover, a good separation of the
different types of fresh and aged oil varnishes is obtained. Thus, resin
identification in oil varnishes is possible through the statistical compar-
ison with a data bank even if the resin absorption covered the oil one.
In fact, resin infrared fingerprint is better conserved in oil varnishes
which lead to better identification via PCA. That is why a specific PCA
was done for the differentiation of different alcoholic and spirit var-
nishes (cf. Fig. 8b). We can see that the different varnish signals are
less separated than the oil one, maybe due to the high standardization
of the signal. Moreover, identification criteria previously given in this
article are sometimes based on weak absorption band, which explain
the results of the statistical analysis.
The Principal Component Spectra are given in Fig. 9. These figures
clearly show that the C\H absorption bands between 2700 and
3100 cm−1 and the carbonyl band account for the most variance
(75.9% of the total variance) in the variables. Thus they are the most use-
ful to identify the different varnish resins and the region of study can be
reduce to 2765–3110 cm−1 and 1550–1900 cm−1. In this way, PCA
made with such region of the spectrum give very similar results than
Fig. 8b (data not shown). Moreover, on the basis of ageing experiment,
we can conclude that these characteristic absorption bands are still
discriminant after 800 h of irradiation in Suntest apparatus. We can
also see that the 2530–2650 cm−1 zone is still discriminant, certainly
because this band is characteristic of colophony. Some of the few
bands appearing in the PCS of the fingerprint region were described
before: 1279 cm−1 (colophony/rosin), 1412 cm−1 (diterpenic resins)
and 1235 cm−1 (fresh diterpenic resins, specific shape when aged).
These results show that the fingerprint zone is not the only pertinent
Fig. 10. FT-IR spectra of the sample from Portrait du Baron Portal.
region to discriminate varnishes of different compositions and ageing
conditions with PCA. Besides they validate some of our characteristic
bands.
4. Application to the identification of artwork varnish
Varnish was sampled from “Portrait du Baron Portal” by Callande de
Champmartin (1797–1883), part of the Fabre Museum (Montpellier).
Fig. 10 gives the FT-IR spectra of the sample. The bands at 3430,
2929, 2875 and 1700–1740 cm−1 show the presence of resin like
organic material. The shape of the C\H band around 2900 cm−1 is
characteristic of a diterpenic resin with a maximum at 2929 cm−1. The
band at 1650 probably shows the presence of a pine resin. The band at
1178 cm−1 is, as said before, characteristic of the photodegradation
of resins and the weak 722 cm−1 band could come from the presence
of oil.
When put in the PCA this varnish is between the dammar oil varnish
and mastic oil varnish (Fig. 8a). For this reason we thought that the
varnish was an oil varnish with a diterpenic resin from Pinus species
and a triterpenic resin. This conclusion was validated by the GC–MS
that showed the presence of various fatty acids and their degradation
products with sterols, notably sitosterol and stigmasterol. Triterpenes
were also found showing the presence of mastic or dammar and
dehydroabietic acid (DHA) and its derivates (7-oxo-DHA, 15-hydroxy-
DHA etc.) were identified confirming the use of coniferous resin
(or by-product).
Acknowledgements
We are grateful to all the conservators and restorators who gave us
insights about the fabrication and use of varnishes and gave us samples
of ancient artworks to study, particularly the Amoroso-Waldeis work-
shop, Thierry Martel and Isabelle Devergne.
 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149 147

Appendix A. Influence of turpentine spirit Appendix B. Venice turpentine vs colophony

Fresh varnish
Venician Turpentine
colophony spirit varnish

absorbance
absorbance

colophony varnish colophony

4000 3500 3000 2500 2000 1500 1000 500

4000 3500 3000 2500 2000 1500 1000 500 n (cm-1)
wavenumber (cm-1)

Aged varnish

colophony spirit varnish

absorbance

colophony varnish

4000 3500 3000 2500 2000 1500 1000 500

wavenumber (cm-1)
148 C. Azémard et al. / Microchemical Journal 112 (2014) 137–149

Appendix C. Drying effect

Sandarac Manilla copal

0 0,6
0h
0,7
14,47 15 h

0,6

0,5
0,4
absorbance

0,4

0,3
0,2
0,2

0,1

0,0
0,0

4000 3500 3000 2500 2000 1500 1000 500 4000 2000
n (cm-1) n (cm-1)
Colophony Mastic
0,9
0h 0h
0,7
15 h 0,8 5h
0,6
0,7

0,5 0,6
absorbance

absorbance

0,4 0,5

0,4
0,3
0,3
0,2
0,2
0,1
0,1

0,0 0,0

4000 3500 3000 2500 2000 1500 1000 500 4000 3500 3000 2500 2000 1500 1000 500

n (cm-1) n (cm-1)

Dammar
0,35
0h
15 h
0,30

0,25

0,20
absorbance

0,15

0,10

0,05

0,00

-0,05
4000 3500 3000 2500 2000 1500 1000 500

n (cm-1)

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