Microchemical Journal 139 (2018) 250–259

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Microchemical Journal

journal homepage: www.elsevier.com/locate/microc

Characterization of natural degradation of historical Moroccan Jewish

parchments by complementary spectroscopic techniques
Latifa Hajji a,b,⁎, Ghizlane Idrissi Seghrouchni a, Abdelhadi Lhassani b, Mohammed Talbi a, M'hammed El Kouali a,
Mona Latifa Bouamrani a, Samia Yousfi a, Chafia Hajji c, Maria Luisa Carvalho d
a
Laboratory of Analytical Chemistry and Physico-Chemistry of Materials, Faculty of Sciences Ben M'Sik, Hassan II University of Casablanca, Morocco
b
Laboratory of Applied Chemistry, Faculty of Sciences and Techniques, Sidi Mohamed Ben Abdellah University, Fez, Morocco
c
Laboratory of Mechanics, Processes, Energy and Environment, National School of Applied Sciences, Ibn Zohr University, Agadir, Morocco
d
LIBPhys-UNL, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Portugal

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

Article history: In this work, we have explored the potentiality of energy dispersive X-ray fluorescence spectrometry (EDXRF), X-
Received 30 September 2017 ray diffraction (XRD), the vibrational attenuated total reflection - Fourier transform infrared spectroscopy (ATR-
Received in revised form 19 February 2018 FTIR), and scanning electron microscopy coupled to energy dispersive X-ray spectrometry (SEM-EDS) for the
Accepted 2 March 2018
characterization of the natural degradation of old parchment. The samples used in this work comprise four Mo-
Available online 3 March 2018
roccan Jewish historical parchments belonging to private libraries. The results have been correlated with those
Keywords:
obtained by a selected modern parchment. The results obtained allowed for the first time an accurate insight
Parchment into the chemical composition of these parchments. EDXRF Analysis of the samples allowed us to study some
Degradation of the undertaken process along the preparation of parchments, such as the liming treatment using CaCO3.
EDXRF XRD was used to elucidate the collagen main features, to identify the inorganic composition of the parchments,
XRD and to evaluate the crystallinity changes upon natural weathering. FTIR spectroscopy enabled us to determine the
ATR spectroscopy changes of collagen material in response to natural ageing, overall oxidation (detection of bands at 1716 cm−1)
SEM-EDS and collagen gelatinization. SEM-EDS results disclosed the morphological surface changes occurring at different
levels in the fibrous network of collagen, and have proved a poor conservation state in some parchments.
© 2018 Elsevier B.V. All rights reserved.

1. Introduction
Parchment is a collagen based material produced from animal skin
such as calves, sheep and goats. Succeeding the papyrus, parchment ma-
terial has been used as a writing support for centuries until it was in turn
dethroned by the advent of papermaking [1]. Historic parchments were
used for several purposes as they preserve in the form of book materials
(manuscripts and scrolls), archival documents, fine art which includes
drawings, paintings and prints; and ritual or sacred objects made for
use as part of religious observance (in Judaism, they include for instance
Torah and Biblical scrolls) [2].
Parchment medium is a complex natural biomaterial and it is impor-
tant to understand and to study its structure. On the other hand, the va-
riety of the manufacturing process traditions and the intrinsic variability
of the animal skin used makes rather difficult the investigation of its
conservation state [1,3]. Generally, its production involves the dehairing
of the skin in lime, washing in water and then thinning and drying
⁎ Corresponding author at: Laboratory of Analytical Chemistry and Physico-Chemistry
of Materials, Faculty of Sciences Ben M'Sik, Hassan II University of Casablanca, Morocco.
E-mail address: lat.hajji@gmail.com (L. Hajji).
under tension on a frame [4]. However, there is no common detailed de-
scription of skins' preparation, as it has been carried out in different
ways depending on the geographical area and historical period in
which the parchment was produced [1]. Therefore, each artifact may
be considered as a unique specimen, and the investigation of its degra-
dation state should be undertaken taken into consideration its own
characteristics.
During the manufacture of parchment, animal skin undergoes a se-
ries of procedures giving rise to a final product composed only on the
dermal skin layer, which predominantly contains heterofibrils of type I
collagen molecules [5].
From the chemical point of view, collagen protein is principally
constituted of three amino-acids (~33% glycine, 20–30% of proline and
hydroxyproline). The type I collagen molecule is a triple helix resulting
from right-handed coiling around the common axis of three α- chains,
each forming a left-handed helix of about 1000 amino acid residues,
with an increase of 0.29 nm per residue in the direction of the
axis. These chains are formed of repeating Gly-X-Y sequences, where
Gly is glycine, X and Y are often proline (Pro) and hydroxyproline
(Hyp), respectively. The triple helix is about 300 nm long and has a
diameter ranging from 1.2 to 1.5 nm [6,7]. The triple helix structure

https://doi.org/10.1016/j.microc.2018.03.006
0026-265X/© 2018 Elsevier B.V. All rights reserved.
 L. Hajji et al. / Microchemical Journal 139 (2018) 250–259
formed by the three polypeptide chains is stabilized by inter-chain hy-
drogen bonds, van der Waals interactions and structural water mole-
cules [5].
The structure of collagen imparts an outstanding resilience and ex-
cellent mechanical and chemical properties to parchment. Some exam-
ples have proved great longevity, such as the scrolls of Khirbet Qumran
caves (Dead Sea Scrolls), those found in Avroman (Kurdistan) and Dura-
Europos (Syria), and which are considered as the oldest parchments
dating back to the 2nd century BCE [8]. Despite its intrinsic structural
strength and resistance, parchment material is subjected to degradation
induced by environmental factors (relative humidity, temperature,
light, pollutants), biological agents (bacteria, fungi, insects, molds), in-
teraction between ink or binders and collagen, and undesirable effects
of the protocols that rules conservation and restoration practices in
use [8]. All these factors can promote different types of deterioration
which alter parchment integrity during time. Oxidation, hydrolysis
and gelatinization are the main mechanisms of its deterioration [9].
Parchment based artworks are among the most precious objects of
Moroccan cultural heritage. There exist a huge variety of historical ob-
jects made of parchments (manuscripts, scrolls and book covers) that
belong to public and private libraries, archives, museums and different
religious foundations. They represent repositories of tangible-intangible
Moroccan cultural patrimony and it is vital that these artifacts remain
well preserved. Moroccan conservators-restorers have long been con-
cerned with the damage induced in parchment manuscripts. Currently,
they are evaluating the conservation state of these documents only in
terms of their visual appearance (color, texture…), but degradation
takes place at several levels and damage cannot always be assessed by
virtue of naked eye.
Enhancing this part of cultural heritage and contributing to its safe-
guard is the reason why we would like to focus on this problem, with
the aim to provide a multidisciplinary approach to understanding colla-
gen degradation, using a variety of techniques capable of detailed anal-
ysis of various levels of the structural hierarchy, capable to produce a
final damage picture.
In the present article, the qualitative aspects of the damage induced
in old Moroccan Jewish parchments were investigated using multi ana-
lytical techniques. The study was conducted on a set of 4 Moroccan Jew-
ish historical parchments owned by different private Moroccan
libraries.
The main objective of this study is to provide a differentiation be-
tween the samples in terms of their chemical composition, the
manufacturing process and to undertake the assessment of the degrada-
tion at macroscopic and microscopic level. Besides, the results obtained
from old parchments have been compared to those of a modern one in
order to better study their conservation state and to monitor the modi-
fications induced by natural ageing.
The main techniques used are; EDXRF analysis which was of great
help in ascertaining the elemental characterization of the samples,
XRD allowed a molecular characterization of the collagen fibers, ATR-
FTIR spectroscopy which enabled the structural characterization and
the study of the degradation pathways, and SEM-EDS that is used for
morphological characterization in order to assess the conservation
state of the samples and to follow the degradation progress within the
collagen fibers network.
The obtained results are of great importance for restorer's commu-
nity in order to better preserve this material in the future and to im-
prove its conservation procedure.
2. Materials and methods
2.1. Sampling
The samples used in this work comprise 4 Moroccan Jewish histori-
cal parchments belonging to private libraries (Fig. 1). The four docu-
ments have different format depending on the size of the related
251
folios or scrolls. Samples V1, V2 and V2T are taken from a manuscript
with folios in the size of 145 × 170 mm. Samples G1 and G2 belong to
a scroll with dimensions of 155 × 400 mm. the I1 and I2 fragments are
from a manuscript of 120 × 150 mm. Samples H1, H2 and H3 are
taken from the borders of a scroll with a size: 290 × 520 mm. However,
all the studied sample fragments have the same similar size: 10
× 10 mm. The results have been correlated with those obtained by se-
lected model samples including a modern parchment that is made
from goatskin. It was produced in the old medina of Marrakech city by
the general traditional process, which involves drying the green skin
under tension, at ordinary temperatures on the sun, brine (NaCl), to
prepare the skin and help removing hair, mechanical scraping with a
proper knife to remove the hair, lime solution (CaO) to soften and re-
move the epidermal layer, polishing the surface with pumice stone
(SiO2·AlO), whitening and opacifying with chalk (CaCO3) and/or gyp-
sum or (CaSO4).
The description of the different samples is presented in Table 1. All
the parchment objects are written in Hebrew. The content consists on
the Torah sacred Jewish religious text and they typically contain
media on only one side. The sampling was made in the ink free area
from the borders as pointed out in Fig. 1 without any kind of sample
preparation. On the other hand, we have analyzed one inked parchment
(V2T) as clarified in Table 1.
2.2. ATR-FTIR spectroscopy
ATR-FTIR measurements were performed with a spectrometer
Bruker Vertex 70 model equipped with a diamond crystal.
The analyses were carried out at room temperature and ambient hu-
midity. All the spectra were acquired between 4000 and 400 cm−1 with
a spectral resolution of 4 cm−1 and 16 scans in order to exploit the in-
strumental built-up noise reduction algorithm. All sample spectra
were captured as transmittance spectra.
2.3. X-ray diffraction
The X-Ray diffraction (XRD) experiments were performed on an X-
Ray diffractometer (X'Pert Pro model) operating with CuKα radiation
(λ = 1.5406 A°). The current (in mA) and voltage (in kV) were adjusted.
The current was adjusted to 30 mA. The voltage was increased to 40 kV.
The reflection angle 2θ is in a range between 10° and 70°, it changes of
0.016° with a step of 40 s.
2.4. Energy dispersive X-ray fluorescence (EDXRF)
The spectrometer used in this work for EDXRF consists of an X-ray
tube equipped with a changeable secondary target, in molybdenum.
With this arrangement, it is possible to obtain a monochromatic source,
which makes possible to select the secondary target in order to get the
best ionization conditions for a special sample. The X-ray tube, the
secondary target, and the sample are in a triaxial geometry. With this
arrangement, we decrease the background, taking the advantage of
the effect of the polarization of the incident X-ray beam from the tube,
and so, improving the detection limits. The characteristic radiation
emitted by the elements present in the sample is detected by a Si(Li)
detector, with a 30 mm2 active area and 8 μm beryllium window. The
energy resolution is 135 eV at 6.4 keV and the acquisition system is a
Nucleus PCA card. Quantitative calculations are made through the
fundamental parameters method. The X-ray generator was operated
at 50 kV and 20 mA and a typical acquisition time of 1000 s was used.
A collimator of silver was placed in front of the detector in order to
restrict the effective area of the detector by excluding regions close to
the edges.
252 L. Hajji et al. / Microchemical Journal 139 (2018) 250–259

Fig. 1. Four ancient Moroccan Jewish parchments in the form of scrolls and pages. (a) parchment H with highlighted areas analyzed (H1, H2 and H3), (b) parchment G (including the
samples G1 and G2), (c) parchment I (with analyzed areas I1 and I2) and (d) parchment V (including V1, V2 and V2T).

2.5. Scanning electron microscopy-energy dispersive X-ray spectrometry
(SEM-EDS)
Environmental scanning electron microscopy scans were obtained
using a Quanta 200 MK2 associated with an EDS microprobe. This mi-
croscope is a high resolution imaging technique that does not require
any sample coating or preparation. SEM observations were made at 5
to 20 kV accelerating voltage; in a pressure of 130 Pa. SEM images
have been obtained using the secondary electron detector at different
magnifications. An energy dispersive X-ray fluorescence microanalysis
system allowed us to obtain elemental analysis. SEM allowed the parch-
ment samples to be observed without alteration.

Table 1
Parchment samples description.

Samples Type of the skin the parchment made of Date Geographical location

V1, V2 Gazelle, written, folios 19th century Meknes

V2T Gazelle, written, folios, inked area 19th century Meknes
H1, H2, H3 Calf, written, scroll 19th century Meknes
G1, G2 Gazelle, written, scroll 19th century Meknes
I1, I2 Unknown, folios 18th century Essaouira
R1, R2, R3 Goat, scroll Modern Marrakech
 L. Hajji et al. / Microchemical Journal 139 (2018) 250–259 253

3. Results and discussions

23,914 ± 1913

20,000 ± 1600

36,000 ± 2880
83,000 ± 6640

1560 ± 125
2850 ± 228
1400 ± 112
540 ± 43

460 ± 37

880 ± 70
3.1. EDXRF

11 ± 1

17 ± 1
V2T
We analyzed the elemental composition of the different parchment
samples by Energy dispersive X-ray fluorescence spectrometry

21,513 ± 1721

37,000 ± 2960
84,000 ± 6720

17,700 ± 1416
(EDXRF). The major advantage of this technique is that it preserves

3540 ± 283
1400 ± 112

1000 ± 80
520 ± 42

700 ± 56

780 ± 62
the integrity of the sample and allows the characterization of each

16 ± 1

11 ± 1
type of parchment in order to understand the manufacturing process

V2
used to produce these artworks.
The main elemental concentrations for S, Cl, K, Ca, Ti, Mn, Fe, Ni, Cu,

24,438 ± 1955

49,000 ± 3920
85,000 ± 6800

22,700 ± 1816
Zn, As, Sr and Pb in the several analyzed parchments are reported in

7100 ± 568
3700 ± 296
1500 ± 120
Table 2. We have analyzed one inked parchment sample (V2T) in

530 ± 42

400 ± 32

910 ± 73

13 ± 1
11 ± 1
order to investigate the ink composition.

V1
The results indicate that within the different parchments Ca, S, CL
and K are the major constituents present, along with several trace

6300 ± 504

4570 ± 366
2300 ± 184
elements.

480 ± 38

107 ± 9
As one observes (Table 2), Calcium exhibits high concentrations

14 ± 1
varying between 2850 ± 228 μg/g in the case of sample V2T and

R3
49,000 ± 3920 μg/g in sample G1. One possible interpretation for the
dominance of calcium is the application of the liming for the skin treat-

4800 ± 384

3200 ± 256
1500 ± 120
370 ± 30
ment during the preparation of parchment. In fact, the liming was ap-

55 ± 4

11 ± 1
plied using a solution prepared based on natural limestone consisting

R2
of CaCO3 [10]. The limestone is transformed to quicklime (CaO) by
heating it to a temperature of 900 °C. After heating, it was ground to a

3500 ± 280

3700 ± 296
2100 ± 168
powder and reacted with water, which imparts an alkaline lime bath.

480 ± 38
The principal role of this bath is to dissolve the epidermal layer of the

95 ± 8

12 ± 1
skin and to soften the fibrous structure by opening the fibers of the der-

R1
mis in order to allow the expansion of the follicles and the hair shaft re-
moval without leaving color residues or blood traces [10]. Thus, the

18,000 ± 1440

10,000 ± 800
2000 ± 160

3300 ± 264
presence of Ca is not surprising since it originates from the liming pro-

510 ± 41
cess (CaO, CaCO3, Ca (OH)2). Chalk (CaCO3) was also employed to give

84 ± 7

27 ± 2

16 ± 1
25 ± 2
52 ± 4
31 ± 3
parchment a whiter surface [11]. Moreover, Ca may be associated
H3

with S due to the sulfur uptake from the atmosphere, giving rise to
CaSO4 resulting from the reaction of sulfur acids with alkaline lime
22,000 ± 1760

[11]. The presence of Calcium corroborates with XRD and SEM-EDS

1650 ± 132

2500 ± 200
9700 ± 776

290 ± 23
results. 41 ± 3

19 ± 2
19 ± 2

25 ± 2
23 ± 2
36 ± 3
It is evident from the results that S and Cl concentrations are among
H2

the highest ones. Indeed, Sulfur content ranges between 1800 ± 144 μg/
g in sample H1 and 85,000 ± 6800 μg/g in sample V1. Concerning Chlo-
22,000 ± 1760

rine, it occurs at concentrations fluctuating in the interval [1400 ± 112

1800 ± 144

2300 ± 184
9400 ± 752

μg/g-24,000 ± 1920 μg/g]. When we consider both K and Cl associated

215 ± 17
45 ± 4

19 ± 2

21 ± 2
24 ± 2
72 ± 6
20 ± 2

to S, they may have been originated from salts used in the surface coat-
H1

ings of parchment, taking into consideration that KCl and KSO4 were
used as preservative in the drying conservation process of the animal
Mean elemental concentration (μg/g) of the historical and modern parchment.

24,000 ± 1920

skins [11].
5500 ± 440
7000 ± 560
4200 ± 336

It was also found that iron occurs at low concentrations with the ex-
125 ± 10

211 ± 17

ception of parchment sample V, where Fe is present at considerably

13 ± 1

11 ± 1

68 ± 5
25 ± 2
9±1

high concentrations probably coming from the ink. Lead exists in values
G2

of the order of 0.2% in only one sample (G1).

Unexpectedly, huge amounts of Zn of the order of 2% and 3% of Ba
24,000 ± 1920

49,000 ± 3920
2400 ± 192

5800 ± 464

2300 ± 184

were detected in the historical samples V1, V2 and V2T. These elements
170 ± 14

127 ± 10

are probably originated from the parchment bleaching process, or as

30 ± 2

14 ± 1
14 ± 1

83 ± 7
35 ± 3

filler used to give the parchment a smooth surface.

G1

3.2. XRD results

3900 ± 312
1500 ± 120
1600 ± 128
4000 ± 320
148 ± 12

255 ± 20

330 ± 26
17 ± 1

23 ± 2
99 ± 8
10 ± 1

The surface of each sheet of parchment was analyzed using X-ray

diffraction (XRD). This analytical method is highly suitable for
I2

complementing XRF in what concerns inorganic matter. Besides, XRD

3900 ± 400
2300 ± 184
1900 ± 152
3400 ± 272

was used for describing the structural hierarchy and molecular packing
345 ± 28

of collagen and for assessing the changes which may occur upon natural
57 ± 5

24 ± 2
49 ± 4

17 ± 1
28 ± 2
12 ± 1

ageing considering that the knowledge of the deterioration of collagen

I1

in parchment and the influence of the degradation factors in its proper-

Table 2

ties are the key to defining the sustainable conditions in which parch-
Mn

Cu
Zn

Pb
Ca

Ba
As
Fe

Sr
Cl

Ti
K
S

ment documents should be stored.

254 L. Hajji et al. / Microchemical Journal 139 (2018) 250–259
Fig. 2a shows the XRD diffractograms obtained for the different an-
cient parchments (I1, H3 and G2) overlapped with the modern one
(R2). All investigated collagen based materials exhibit XRD pattern
characteristic of collagen, in which the main diffraction peaks appear
very distinctly at the diffraction angle 2θ values of 2θ = 7.8° (peak 1),
2θ = 17.3° (peak 2) and 2θ = 40.4° (peak 3). The first peak is sharp
and is related to the highly ordered region of the collagen fibrils. It re-
flects the distance between one collagen molecule and its nearest neigh-
bor or diamater of the thri-helix collagen molecule [12]. The second
wide peak presented in a diffuse scattering is characteristic of the amor-
phous region that is less ordered [12,13]. The third crystalline peak is a
small scattering and is referable to the helical rise residues or the dis-
tance between the amino acid residues along the collagen molecular tri-
ple helices [12,13].
On the other side, XRD analysis of the samples has revealed struc-
tural changes to the molecular packing of collagen. As it can be seen in
Fig. 2a, high differences among the intensities of the peaks were ob-
served. We can notice that the peak intensities for the new parchment
are higher than the old ones, meaning that the loss of the structural in-
tegrity is principally linked to the ageing process. Also noteworthy is
Fig. 2. XRD diffractograms corresponding to: (a) old parchment samples I1, H3 and G2 overlapped with the modern one R2, (b) old parchment V2.
that sample H3 is the most degraded among all the samples because
the first peak related to the highly ordered region has completely
disappeared.
Regarding the inorganic composition of the parchment, XRD pat-
terns have revealed the presence of calcium carbonate in some samples
(ancient parchment G2 and H3). The presence of CaCO3 in XRD
diffractograms is interpreted as resulting from the manufacturing pro-
cess, either from surface treatments with chalk or immersion in slaked
lime [14]. Sample V2 (Fig. 2b) presents the typical characteristic peaks
of barium sulfate as a filler added during the manufacturing process.
This founding is in agreement with EDXRF results.
3.3. ATR-FTIR results
Attenuated total reflectance infrared spectroscopy was applied to
parchment samples for the qualitative analysis of such complex mate-
rials with the aim of characterizing their chemical structure. ATR-FTIR
was also useful to identify the natural ageing-induced changes at the
molecular level of collagen, by comparing the spectra of old parchments
with the modern one.
 L. Hajji et al. / Microchemical Journal 139 (2018) 250–259 255

Fig. 3. Overlay of ATR-FTIR spectra of old historical parchments (V2, I1, H2 and G1) and a modern sample R3. Spectrum details: a) region between 4000 and 400 cm−1, b) region between
2000 and 400 cm−1.

Representative IR spectrums of old historical parchment samples V2, band located in the interval of 3437–3414 cm−1 corresponding to
I1, H2 and G1 were compared with a new parchment R3. The spectra the stretching vibrations of both OH and NH variously hydrogen
were recorded in the frequency regions 4000–400 cm−1 and 2000– bonded [3].
400 cm−1 as shown in Fig. 3a and b. The summary of IR vibrational as-
signments was done in accordance with the literature [15–21] and can
be found in Table 3. Table 3
Assignments of IR bands for the investigated parchment samples [15–23].
The spectra of all parchments (Fig. 3a) display typical bands related
to the peptide linkages of type I collagen. The main features are Amide A Wavenumbers Description of vibrational mode
characterized by a broad band at 3400–3100 cm−1 assigned to first (cm−1)

component of Fermi resonance between N\\H stretch and the overtone 3437-3414 ν(OH, NH)
of Amide II, Amide B band (3080–3070 cm−1) ascribable to the second 3400-3100 Amide A: first component of ν(NH) in Fermi resonance with the
amide II overtone
component of Fermi resonance between N\\H stretch and the overtone
3080-3070 Amide B: second component of ν(NH) in Fermi resonance with
of Amide II. Another prominent band is Amide I absorption band rang- the amide II overtone
ing in the interval 1643–1630 cm−1 and that is attributed to C_O 2956-2857 ν (CH3) asymmetric
stretching vibrations of peptide bonds. We can observe the Amide II 2920 ν (CH2) asymmetric
band at 1550–1532 cm−1 (corresponding to CN stretching and NH 2857 ν (CH3, CH2) symmetric
1716 ν (C = O)
bending vibrations). The Amid III absorbance near 1240–1235 cm−1
1643-1630 Amide I, ν (C = O)
arises from NH in plane bending and CH2 wagging vibration of glycine 1550-1532 Amide II, ν (CN) and δ(NH)
backbone and proline side chain [15–21]. 1450-1415 ν (CO2−
3 ) in CaCO3

Parchment is mainly made from collagen and type I collagen is δ(CH2) and δ (CH3)hidden by CaCO3
1240-1235 Amide III, in plane δ (NH) and CH2 wagging
principally composed of three amino acids (∼33% glycine, 20–30%
1046 ν(C\\O) or skeletal stretching, found in proline and
proline and hydroxyproline). Its structural unit is a triple-helix; heli- hydroxyproline.
ces are arranged to form fibrils and, at an upper hierarchical level, fi- 1023 ν(C\\N) in glycine, proline and hydroxyproline
brils compose the final collagen fibers. Since the triple helices of 720 δ (OCO) in CO2–3 groups (in-plane deformation) of CaCO3 and

collagen are held together by hydrogen bonds, we have also detected CH2 rocking

the presence of another band overlapping with amid A and B. It is the * ν stretching; δ bending.
256 L. Hajji et al. / Microchemical Journal 139 (2018) 250–259
Parchment band at around 1023 cm−1, clearly visible in the spectra
of sample G1, is interpretable as C\\N stretching in glycine, proline and
hydroxyproline. Another band located at about 1046 cm−1 is found in
proline and hydroxyproline and can be attributed to C\\O stretching
or skeletal stretching [22].
Moreover, additional information about the inorganic composition
of parchment can be obtained. Indeed, the presence of carbonate
(mainly CaCO3) is confirmed by the band at around 1450–1415 cm−1
assigned to the asymmetric C\\O stretching mode and another weak
peak at 720 cm−1 ascribable to O-C-O in plane bending in CO2– 3 groups
[19,23].
FTIR spectra were of great assistance in examining the process of
degradation of the analyzed samples. In general, parchment alteration
is due to collagen degradation that is the result of denaturation, hydro-
lysis and oxidation. The comparison between the spectra of ancient
parchments with the modern one enabled us to examine the positions
and intensities of some bands important to evaluate the molecular alter-
ation of collagen. The principal conclusions drawn from this comparison
are:
• The decrease of the intensities of the bands of Amide A, Amide B and
hydrogen bonded water that are located in the interval 3450–
3100 cm−1 (Fig. 3a). This decrease is highlighted in the case of sample
I1 and particularly emphasized in the spectra of old parchment V2;
which indicates the destruction of hydrogen bonds formed between
Fig. 4. SEM micrographs under different magnifications (500×, 1000×, 1500×) using LFD images of ancient parchment samples G2, H1, I1, V1, V2T and a modern parchment R3.
the triple helix of the collagen molecule and therefore, the partial de-
struction of the collagen molecule into a random coil upon natural
ageing [25].
• The formation of a new shoulder at 1716 cm−1 in the case of ancient
parchment sample H2 (Fig. 3b), which corresponds to the formation
of carbonyl and carboxylic groups, suggesting the oxidation of colla-
gen molecules [15]. This band may also be attributed to tannin com-
ponents, considering that in Jewish parchments, different
manufacturing traditions had been historically applied, involving the
use of tanning compounds on the writing support surface, used with
alum and other salts, according to some ritual procedures. However,
spectroscopic evidence of the tanning treatment using only FTIR spec-
tra is not sufficient, and should be normally confirmed by another an-
alytical technique such as Raman spectroscopy.
• The oxidation of lateral amino-acid side chains like methionine, sug-
gested by the presence of a new band in the range 1170–1150 cm−1
(Fig. 3b) in the case of samples V2 and H2 [15].
• The decrease in the intensity of the bands related to amide I and II
(Fig. 3b) in what concerns historical parchments G1, I1 and V2. This
indicates the denaturation of collagen to gelatin or the conversion of
the triple helix to disordered structures.
• The Increase of the absorbance at 1407 cm−1 (primary amines) in the
case of parchment G1 (Fig. 3b), which is suggestive of the scission of
peptide bonds and the hydrolytic degradation of collagen structure
[26,27].
 L. Hajji et al. / Microchemical Journal 139 (2018) 250–259
• The reduction of the contribution of CaCO3 content indicated by the
decrease of the intensity of the band at 1433 cm−1, clearly visible in
parchment V2.
3.4. SEM-EDS
Microscale morphological examination was made by environmental
scanning electron microscopy which does not require sample coating. In
order to characterize the surface morphological variations in response
to natural ageing, SEM pictures of aged and modern parchment samples
were recorder at several magnifications ranging from 500× to 1500× as
shown in Fig. 4. Besides, Observations performed by scanning electron
microscopy magnify the typical parchment morphology and the struc-
ture of collagen fibers, emphasizing the conservation state of the sam-
ples. Energy dispersive spectrometry allows the determination of the
elemental composition of the inorganic compounds added during the
fabrication of the parchments.
The micrographs obtained in the secondary electron mode show a
heterogeneous fibrous structure as fibers have different shapes, sizes
and orientations. It is important to note significant aspects of degrada-
tion manifested by the presence of massively shrunk fibers as it is the
case of sample I1 (Fig. 4), for which the surface morphology exhibits
vulnerable conservation state. Its surface shows very fragmented and
detached layers with the presence of swollen fibers. This confirms the
damage of collagen as it was also evaluated by FTIR spectroscopy,
whose results proved the occurrence of collagen gelatinization in this
sample. Noteworthy is the detection of a glossy surface as another as-
pect of deterioration, as it can be seen in the micrograph of sample H1
(Fig.4).
The investigation of the modern parchment R3 evidence a good con-
servation state manifested by a densely packed collagen fibrous net-
work, whereas a poor conservation state was observed for parchment
V1 for which the fibrous network is not recognizable. Exclusively with
Fig. 5. SEM micrographs of sample V2T under 1500× magnifications (a) with EDS spectra (b) supported by quantification results.
257
parchment sample V, SEM micrographs have been performed on the
inked area (sample V2T), and it can be seen that the morphological fea-
tures of the fibrous network are not visible. SEM image of parchment
sample V2T (Fig. 5a) depicts bright particles heterogeneously spread
on the surface of the material, in which EDS elemental analysis
(Fig. 5b) identifies as Ba, Fe, Zn, Pb and S present in important levels.
The presence of Ba associated with that of S is probably coming from
BaSO4, which is supported by XRD results. Fe originates from ink resi-
dues. The detection of Zn in relevant amounts exclusively in this sample
perfectly corroborates with EDXRF analysis. Other trace elements were
also detected, such as Al and K. The presence of these 2 elements
coupled to that of S, may be linked to the addition of alum (KAlSO4) as
a tawing agent [28].
A further closer look at the micrographs of sample H1 (Fig. 6) has
shown that the collagen fibrous network still persistent despite the deg-
radation aspects detected. We can observe melt like fibers highlighted
in the form of a glossy surface (Fig. 6a), but the presence of an undam-
aged well conserved fiber next to it suggests that degradation is not
progressed on the whole surface of the parchment. Fig. 6b emphasizes
the glossy part showing the fusion of the broken fibers that may be cor-
related with the gelatinization of collagen [15].
Energy dispersive X ray spectrometry was also performed in some
cases, in order to ascertain the elemental composition of the inorganic
compounds. Fig. 6c displays the results obtained for parchment sample
H1 and which are in a very good agreement with EDXRF; the vestiges of
manufacturing process are evidenced mainly by the detection of Ca, Cl,
K and S originating from the liming applied for the dehairing of the skin
and the drying process of parchment.
4. Conclusions
Our research allowed for the first time an accurate insight into Mo-
roccan historical Jewish parchment, especially in what concerns the in-
vestigation of the organic material, the inorganic fillers added during
258 L. Hajji et al. / Microchemical Journal 139 (2018) 250–259
Fig. 6. SEM micrographs of ancient parchment sample H1 with the corresponding EDS
elemental analysis: (a) 500×, (b) 1000×, and (c) EDS spectra with quantification results.
the manufacturing process and the assessment of the degradation path-
ways of collagen from molecular, structural to macroscopic levels.
The use of a multi-analytical approach based on four complementary
analytical techniques (EDXRF, XRD, ATR-FTIR and SEM-EDS) was suc-
cessful to assess the conservation state of the different ancient samples
by making a comparison with the results obtained from a modern
parchment used as a reference.
EDRXF proved again to be of great usefulness to ascertain the ele-
mental composition thanks to its capacity to detect major and minor el-
ements, permitting the characterization of the material and the study of
the manufacturing process of these parchments. XRD was used as a
complementary technique that enabled the study of the structural
changes as the collagen lattices degraded upon ageing.
This work also demonstrated that ATR-FTIR spectroscopy is a highly
reproducible technique that is important for analysis of historical parch-
ments, as it offers valuable information on the chemical changes of col-
lagen in response to weathering. SEM-EDS disclosed different aspects of
degradation in the morphological surface of parchments, underlying the
poor conservation state of some samples and evidencing the changes af-
fecting the collagen fiber network.
In conclusion, our findings pointed to describe a relative damage pic-
ture of these parchments and to discriminate between them in terms of
organic and inorganic composition. The results prove once again that a
multi analytical approach is indispensable in the analysis of cultural her-
itage artifacts, for their preservation and also to improve the restoration
procedures.
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