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Analytical techniques used for the evaluation of a 19th century
quranic manuscript conditions
Gomaa Abdel-Maksoud ⇑
Conservation Department, Faculty of Archaeology, Cairo University, Giza, Egypt
a r t i c l e i n f o
Article history:
Received 14 June 2010
Received in revised form 10 May 2011
Accepted 21 June 2011
Available online 29 June 2011
Keywords:
Paper
Leather
Microscopes
XRD
EDAX
Microorganisms
Chemical analysis
HPLC
a b s t r a c t
The manuscript studied here dates to 19th century, and consists of paper pages and leather
bookbinding. This study aims to use analytical techniques in order to identify the compo-
nents of the manuscript and to explain its deterioration process. Visual assessment, isola-
tion and identification of fungi, pH measurements, and investigation of the surface
morphology by a scanning electron microscope (SEM) were used to explain paper and
leather deterioration. X-ray diffraction with EDAX, Fourier Transform Infrared Spectros-
copy (FTIR), and chemical analysis were used to identify pigments, binder of pigments,
ash, lignin, and the a-cellulose content of papers. The shrinkage temperature measurement
was used to explain the deterioration process of leather. SEM was used to identify the type
of animal skin used for the bookbinding and high performance liquid chromatography
(HPLC) was used to identify the vegetable tanning material used with the bookbinding.
The results revealed that the ink used was a mixture of carbon with iron gall. The pig-
ments used on the paper were gold leaf or gold shell, cobalt oxide, and mercuric sulfide
for the gold, blue and red colors respectively. Sodium chloride was the main salt crystal-
lized on the surface of paper. Calcium carbonate was the filler used in the paper making
process. Cotton fibers may have been used as a raw material in the creation of paper.
The values of the shrinkage temperature and pH were lower than in normal conditions,
indicating that the leather bookbinding suffers from deterioration. Aspergillus sp., and
Penicillium sp. were the most dominant fungi found on the manuscript. Goat skin was
identified as the animal skin of the bookbinding, and Acacia Arabica was identified the
tanning material used with the bookbinding. The condition of the manuscript studied with
its components play an important role in its deterioration.
2011 Elsevier Ltd. All rights reserved.
1. Introduction
A major problem in the preservation of cultural heritageis the damage to manuscripts, mainly paper and bookbind-
ing. Damage can be caused by the effect of aggressive
atmospheres, humidity and temperature, alterations of
paper constituents, and added materials such as ink and
hand coloring with pigments or dyes [1]. Deterioration of
paper-based materials is mainly due to the degradation
of cellulose caused by a number of factors, such as chemi-
cal attack due to acidic hydrolysis, oxidative agents, light,
air pollution, and biological attack due to the presence of
microorganisms like bacteria and fungi [2,3].Leather products, such as the tanned skin of the book-
binding, have been useful materials since the dawn of
human history [4]. Vegetable tanned leather was used in
Egypt throughout the ages starting from the Prehistoric
Period [4] up until the present time [5,6]. Vegetable tanned
leather represents a very complex material composition.
The surroundings of the manuscript are likewise a very
complex and dynamic dimension, constantly varying with
respect to the quantity and degree of their interaction with
each other and with those materials stored within them
[7]. The most common types of damage sustained by a
0263-2241/$ - see front matter 2011 Elsevier Ltd. All rights reserved.doi:10.1016/j.measurement.2011.06.017
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E-mail address: [email protected]
Measurement 44 (2011) 1606–1617
Contents lists available at ScienceDirect
Measurement
j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e/ m e a s u r e m e n t
http://dx.doi.org/10.1016/j.measurement.2011.06.017mailto:[email protected]://dx.doi.org/10.1016/j.measurement.2011.06.017http://www.sciencedirect.com/science/journal/02632241http://www.elsevier.com/locate/measurementhttp://www.elsevier.com/locate/measurementhttp://www.sciencedirect.com/science/journal/02632241http://dx.doi.org/10.1016/j.measurement.2011.06.017mailto:[email protected]://dx.doi.org/10.1016/j.measurement.2011.06.017
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bookbinding are caused by poor handling, poor storage
methods, inappropriate display methods, wear and tear
from repeated use, chemical changes in the materials mak-
ing up the leather objects, chemical changes caused by
atmospheric pollutants and chemicals in contact with the
leather objects, and a combination of any or all of these
(http://archive.amol.org.au) [8].
Chemical deterioration of leather occurs through two
competitive and interactive chemical mechanisms,
oxidation and acidic hydrolysis [9]. Oxidation is generally
caused by free radicals generated by heating, UV light,
and SO2 and NO x pollution. The side chains of some amino
acids are initially involved, but oxidation can also occur in
the backbone of collagen through the rupture of N–C cova-
lent bonds [10]. Hydrolysis is catalyzed by both hydroxyl
and hydrogen ions, especially when atmospheric pollu-
tants such as SO2 and NO x act in conjunction with air
humidity, and cleavage of peptide bonds disrupts the
hierarchical structure of collagen [10]. Gelatinization, in
further denaturation and aggregation, leads to the irrevers-
ible formation of a heavily hydrated gel matrix. Partially
degraded collagen is especially susceptible to gelatiniza-
tion in warm, damp environments, since H-bonds are
exposed to the action of water [10].
The development of specific analytical techniques
improves the procedures to authenticate patrimonial ob-
jects made from collagen and cellulose-based materials
as well as the methods to study the impact of the environ-
mental factors. During the last few decades, many methods
of the analysis of paper and leather have been used for the
identification of the material compounds and for the esti-
mation of the deterioration processes. For paper, Ververis
et al. [11] used chemical analysis for the determination
of hemicelluloses, lignin, and ash content in paper. Strlič
et al. [12] mentioned that the measurement of the pH of
historical paper plays an important role to explain the
mechanism of deterioration. Many authors used different
analytical techniques for the identification of pigment used
on paper manuscripts [1,13,14]. FTIR was used for the
identification of ink binder [15,16]. For the leather book-
binding, the measurement of the shrinkage temperature
is vital and is considered one of the most important tools
used for the determination of leather deterioration [16–
21]. The measurement of leather pH reveals the state of
leather inside a museum or in storage [7,22]. Microbiolog-
ical studies and investigation of the surface morphology
are also very important for the estimation of the deteriora-
tion process for paper and leather [22–24].
This study aims to identify thematerialsused in theman-
uscript studied, apply the most effectiveness techniques of
analyses for the determination of paper and bookbinding
degradation, and explain the mechanism of deterioration.
2. Historical background
The manuscript was found in the library of Ahmed Al-
Bajam Mosque, located in Mehalit Marhoum Village, Tanta
City, Egypt. It was discovered during the destruction of the
mosque. It dates back to 19th century, and contains part of
the 28th chapter of the Holy Qur’an. It suffers from ground
and surrounding environmental damages, especially from
salts, such as sodium chloride.
3. Material and methods
For the determination of materials used with the man-
uscript and to explain the deterioration processes, all the
analytical techniques used in this study (pH, SEM, EDAX,
X-ray diffraction, FTIR, measurement of the shrinkage
temperature, HPLC and chemical analysis of paper) were
considered micro-destructive techniques because a few
micrograms are needed and can be collected from the loose
and separated fibers from the manuscript. The micro-
destructive techniques used were selected to obtain a sig-
nificant identification and to obtain the optimum amount
of information concerning the materials used. The analyti-
cal techniques used were more effective in explaining the
deterioration processes of the manuscript.
3.1. Visual assessment
Visual assessment, by the critical eye of the author, was
applied to determine the aspects of deterioration found on
the manuscript’s paper and leather. This method is very
effective because the causes and mechanism of deteriora-
tion may be easily identifiable. The critical eye of conserva-
tor can also determine the most effectiveness techniques of
analysis, which should be applied for identifying the condi-
tion of the manuscript studied.
3.2. Isolation and identification of fungi
Sterile swabs were used to wipe the surface of the paper
and the leather to isolate the fungi, especially in the
contaminated area. Isolation was made directly in the
laboratory after wiping process. The fungi were isolated by
wiping the swabs on culture medium of potato-dextrose-
agar (PDA) then incubated at 28 C for 1–2 weeks. Czapek
yeast extract agar (CYA) composed of K2HPO4 1 g, Czapek
concentrate 10 mL, yeast extract (Difco) 5 g, agar 15 g, dis-
tilled water 1 L [25]. The source of carbon (sucrose) was
not used andthe paperand leathersamples were thesource
of carbon. Seven-day cultures on Malt Extract Agar (MEA),
which consists of maltose 12.75 g, dextrin 2.75 g, glycerol
2.36 g, peptone 0.78, agar 15 g and distilled water 1 L, was
used for identification of isolated fungi [25]. Fungi colonies
were identified according to Raper and Fennell [26], Barnett
and Hunter [27], and Watanabe [28]. The balance used to
weight the components of media used for the isolation
and identification of fungi was calibrated using standard
weights traceable to SI (International Measurement Sys-
tem). All glass wares used here were calibrated and have
traceability to SI. Isolation and identification of fungi were
carried out at microbiological laboratories, the Department
of Microbiology, Krakow University of Agriculture, Poland.
3.3. pH measurement
3.3.1. Determination of leather bookbinding pH
The pH value of leather was determined by Abdel-Mak-
soud [22] in accordance with Wouters et al. [29] and the
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National Library of the Netherlands [30] but with some
modifications. Samples (0.025 g) of leather bookbinding
is enough for the measurement and was taken mechani-
cally, in the form of loose fibers, from as near as possible
to the damaged area on the surface (grain) of the leather.
The sample was cut into very small pieces. The pH was
measured approximately 6 h after the suspension had been
prepared to allow the ions to migrate into the solution. The
measurement of pH value of the bookbinding was done
using a 315i InhaltsverzeTechnische Werkstatten GMBH
& Co. UG provided with a combination electrode and cali-
brated to between 2 and 7, at 21–22 C. The calibration
of pH meter was done by immersion of the pH electrode
firstly in distilled water and secondly into buffer solutions
(2 and 7). The model numbers of the technical buffers used
was 108,708 for the first buffer (pH 2) and 108,706 for the
second buffer (pH 7). After each measurement of the buf-
fers used, the pH electrode was rinsed in distilled water
followed by immersion in the buffer solutions. The calibra-
tion of pH meter was ok when the reading of pH meter was
stable and closed to the selected buffer value. The mea-
surement was performed in the Central Laboratory, Faculty
of Archaeology and Anthropology, Yarmouk University,
Jordon.
3.3.2. Determination of paper pH
The measurement of the pH of the paper was in accor-
dance with Strlic et al. [31] with little modification. A drop
of distilled water was placed on the paper of the manu-
script, the flat-surface combined pH electrode pressed
against it, and the pH value read after being constant for
30 s. The results are an average of five determinations.
The pH-meter Metrohm 691 (Metrohm, Herisau, Switzer-
land) was used with flat combined electrode (Metrohm
6,0253.100). Before the measurement of pH, the sensor
was calibrated using the provided buffer solutions accord-
ing to the instructions placed nearby the pH meter, pH
measurement was calibrated using three two standard
buffers (Metrohm buffers) of 4 and 7. The electrode with
immersed assembly in the first buffer and the temperature
measured was 25 C when the reading was stable and
closed to the selected buffer (pH 4), the measurement of
the first buffer was finished. The electrode was rinsed
again in distilled water. The pH electrode was rinsed into
the second buffer solution. When the reading of pH was
closed to the second buffer (pH 7), this means that the
pH meter was ready to measure the pH value of the histor-
ical paper samples. The measurement was performed at
the Organic Chemistry Laboratory, Department of Chemis-
try, Krakow University of Agriculture, Poland.
3.4. Investigation of the surface morphology by SEM and EDAX
analysis
A scanning electron microscope, JEOL-JSM-5400LV, was
used for the investigation of the surface morphology of the
paper and the leather. The fine gold coating (JEOL-JFC-
1100E) was used. All samples were photographed by SEM
at the Scanning Electron Microscope Laboratory, The central
Laboratory unit, Assiut University, Egypt. This laboratory
achieves the traceability via the manufacturer of the
instrument throughout the routine maintenance to achieve
best performance. For EDAX analysis, link ISIS Oxford was
used. The quantitative methodused wasZAF. Theresults ob-
tained from EDAX were automatically normalized to 100%.
EDAX was calibrated by the standard nickel rod purchased
along with the instrument. The Calibration was conducted
by count calibration mode in the software. EDAX were also
performed at the Scanning Electron Microscope Laboratory,
The central Laboratory unit, Assiut University, Egypt.
3.5. X-ray diffraction and EDAX analysis of ink and pigments
used
The ink samples of red, blue, and gold colors were
analyzed by X-ray diffraction using Compact X-ray Diffrac-
tometer System PW 1840 – Analytical Equipment – Philips
– Eindhoven – the Netherlands (CU Ka radiation with
Ni-filter). Calibration of X-ray diffraction was done by
using a silicon standard sample. The tube of copper was
used for the measurement. The mA and kV were adjusted.
The mA control was adjusted until the meter indicates at
50 mA. The kV was increased to 40. Two reflection angles
for the silicon standard should appear. One should appear
at 28.44 h and the other appears at 56.12 h. By this opera-
tion, the instrument was ready for the measurement. The
measurement was performed at the Laboratory of X-ray
diffraction analysis, Conservation Department, Faculty of
Archaeology, Cairo University, Egypt. Link ISIS Oxford
was used for EDAX analysis. The results obtained from
EDAX were automatically normalized to 100%. It should
be mentioned that all the decimal points obtained from
EDAX were effective. EDAX were performed at the Scan-
ning Electron Microscope Laboratory, The central Labora-
tory unit, Assiut University, Egypt.
3.6. Identification of pigment binder by FTIR
In order to identify the binder of the ink and pigments
used on paper of the manuscript, a few milligrams of the
ink and pigments taken from the manuscript was ground
into a powder and then mixed with KBr and placed in a
DRIFT cell. The measurement range is between 4000 and
400 cm1. The examination was done by using an infrared
instrument (Bruker) to identify the binder used with black
ink and pigments. Before measurement, the standard sam-
ple provided by the company of instrument is polystyrene
film. According to the instruction of the manufacturer, the
ideal measurement of this sheet must be placed in the
same position and intensity as the standard spectrum
saved at the library of the instrument. Before measurement
process, the background measurement was performed in
order to reduce the effect of the atmospheric carbon diox-
ide and water vapor. FTIR was performed at the Laboratory
of FTIR, Microanalysis Laboratory, Faculty of Science, Cairo
University, Egypt.
3.7. Measurement of hydrothermal stability of leather
bookbinding
This measurement was in accordance with Larsen [7]
but with little modification. A sample of about 0.3 mg fiber
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from the corium part of the leather was wet with distilled
water for at least 10 min on a microscope slide. Then the
samples were transferred to another slide with acetone
for at least 30 min and again transferred to distilled water
and glycerin. The fibers are separated, air bubbles were
removed with a needle, and the fibers were well dispersed
on the slide and secured. The microscope slide was placed
on the hot table. The gradient heat controller was cali-
brated at 22 C, as well as 80 C to achieve the traceability
to ITS 90 (International Temperature Scale). The timer was
calibrated using a calibrated stope watch traceable to
NIST–USA (National Institute for Standard and Technol-
ogy). The measurement of hydrothermal stability was
performed at the Analysis Laboratory, Animal Physiology
Department, Jagiellonian University, Poland.
3.8. Identification of animal skin used for the bookbinding
The surface examination by a scanning electron micro-
scope, JEOL-JSM-5400LV, was used to identify the type of
animal skin used for the leather bookbinding. A specimen
of about 2 mm 4 mm was aligned on a stub, with the hair
follicles opening towards the stub as viewed under the
microscope. The specimen was mounted on stub and
coated with fine gold. The nominal thickness of the sample
was 20 nm. The study of Haines [32] was taken as a refer-
ence in order to be compared with the identified skin from
the bookbinding studied. SEM was performed at the Scan-
ning Electron Microscope Laboratory, The central Labora-
tory unit, Assiut University, Egypt.
3.9. Identification of tanning material by High Performance
Liquid Chromatograpy (HPLC)
The samples and analytical procedure for High Perfor-
mance Liquid Chromatograpy (HPLC) were in accordance
with Wouters [33], and Abdel-Maksoud [34]. HPLC system
was calibrated for the flow rate using calibrated stopwatch
(traceable to standard cesium clock that has direct trace-
ability to SI measurement system at NIST–USA) and volu-
metric flask traceable to Si Measurement system. The
flow rate uncertainty was found 0.02%. Regarding the par-
ticle sizes, the system was calibrated using the standard
polystyrene dispersed particulates in aqueous system of
sizes 2 and 3 lm. Photometric scales was calibrated using
standard [0.1 N] potassium dichromate solution and the
total expanded uncertainty was found ±0.20% with cover-
age factor of two to give confidence level of 95%.
Samples were prepared from the bookbinding and from
Acacia Arabica, supplied by the Commercial Tannery, Cairo,
Egypt, and mimosa and quebracho powders, supplied by
Abd El-Rahman M. Harraz. Agricultural Seeds, Spices and
Medicinal Plants Company, Cairo, Egypt. A sample of dry
vegetable-tanned bookbinding weighing 50 mg was col-
lected from loose fibers around the bookbinding. 100 mg
of each of the new tanning powders was weighed and
soaked in a solution of water/acetone (1/1, v/v) in closed
vessels at room temperature for 24 h. The volume of the
extract liquid was 20 mL for 20 mg of acclimatized vegeta-
ble tanned bookbinding and tanning powder, weighed
after conditioning for at least 48 h at 65%RH and 20 C. Just
before chromatography, an aliquot of the water/acetone
extract was diluted fivefold with methyl alcohol. Extract
analyses were performed immediately. The results pre-
sented here are those recorded for the bookbinding and
Acacia Arabica powder because of their similarity. The
samples were analyzed using a Varian Pro Star, 3 lm,
100 6.4 mm column; pump (Varian Pro Star 230); 990+
photodiode-array detector (Varian Pro Star 335); data
handling using Star chromatography WS (work station)
version 6; 20 lL sample loop; flow rate: 1.2 mL/min; ana-
lytical wavelength 280 nm (other postrun selections possi-
ble between 200 and 800 nm); flow scheme (A = methanol,
B = water, C = 50 g/L phosphoric acid in water): 10A/80B/
10C for 2 min, linear gradient to 90A/10C over 17 min,
90A/10C for 3 min; temperature 21 C. The identification
of tanning was performed at the Analytical chemistry
Laboratory, the Department of Chemistry, Yarmouk
University, Jordan.
3.10. Chemical analysis of paper
3.10.1. Ash content
The ash was estimated by igniting in a muffle furnace a
weighed sample in a porcelain crucible for 30 min at
400 C, then continuously for 45 min at 850 C and then
gravimetrically estimated (Tappi standard 211-om 85)
[35]. The percentage of ash was calculated from:
Ash% ¼ Weight of ashðafter ignitionÞ
Weight of dry paper sampleðbefore ignitionÞ
100
Muffle furnace was calibrated at 400 C, 850 C and
1000 C using calibrated thermocouple that has traceabil-
ity ITS 90 (International Temperature Scale). Ash contentwas determined at the Cellulose Laboratory, Department
of Cellulose and Paper, National Research Center (NRC),
Egypt.
3.10.2. Lignin
An exact weight of 1 g of the air-dried sample was
treated with 15 mL of 72% sulfuric acid for 2 h at room
temperature. The material was then transferred into a 1 L
flask, diluted with 560 mL of distilled water, and boiled un-
der reflux for 4 h. The lignin was filtered on a previously
weighed dry ashless filter paper, and then washed with
hot distilled water till neutrality. The filter paper and lignin
were transferred to a weighted porcelain crucible and
dried in an oven at 105 C (The drying oven was calibrated
at 105 using calibrated glass thermometer traceable to ITS
90) till constant weight.
For ash correction, the contents of the crucible were
ignited at 400 C for 30 min and then at 850 C for further
45 min. The weight of the ash was subtracted to give the
weight of the pure lignin. Lignin percent is calculated from:
Lignin% ¼weight of lignin Weight of its ash
Weight of moisture free pulp 100
Determination of lignin was performed at the Cellulose
Laboratory, Department of Cellulose and Paper, National
Research Center (NRC), Egypt.
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3.10.3. a-cellulose content
25 mL of 17.5% sodium hydroxide solution was added to
3 g, exactly weighed, and cut into small pieces. The slurry
after being left to swell for 4 min, time exactly measured
from the last drop, at 20 C, was pressed for 3 min, with
glass rod, and 25 mL 17.5% sodium hydroxide solution
were added. The suspension was thoroughly mixed for
about 1 min and left covered at 20 C. After 35 min,
100 mL of distilled water was added followed by filtration
in a sintered crucible. The filtrate was poured twice on the
paste before washing with distilled water untill complete
neutrality and then with 10% acetic acid followed by dis-
tilled water. The temperature must be kept constant at
20 C during the whole experiment. The a-cellulose was
estimated gravimetrically after dryness in drying oven at
105 C for 6 h. The percent of a-cellulose is calculated as
follow:
% a-cellulose ¼B
A 100
where A represents the weight of dry sample, and B is theweight of dry treated samples.
All glass wares used for the determination of a-cellu-
lose content a-cellulose content were calibrated and have
traceability to the Egyptian National kilogram number
58. Determination of a-cellulose was performed at the Cel-
lulose Laboratory, Department of Cellulose and Paper, Na-
tional Research Center (NRC), Egypt.
4. Results and discussion
4.1. Visual assessment
The storage of the Qur’an manuscript was very poor andled to advanced deterioration. The following aspects of
deterioration were noticed on the leather bookbinding
(Fig. 1A and C): folding of the fibers in some places, white
hard crust may be from storage, general hardness, fungal
spots, erosion of tanning material, holes caused by insects,
and missing parts. The following aspects of deterioration
were noticed on the papers of the manuscript (Fig. 1B
and D): dog-earred pages, local damage at the corners with
multiple folds and creases, turning over of the paper so
that the front or back surface is in contact with itself, local
missing parts at the corners with multiple folds and
creases, stains derived from different sources (fungi, fats,
dusts, ink and pigments) and improper restoration, totaldestruction of paper edges and sometimes turned to pow-
der, and salt crystallization on the surface of paper and led
to erode the papers.
4.2. Identification of fungi
The results of this study revealed that the most domi-
nant fungi on papers of the manuscript were: Penicillium
restrictum, Penicillium spinulosum, Penicillium rubrum, Peni-
cillium chrysogenum, Aspergillus fumigates, Aspergillus niger,
Aspergillus flavus, Aspergillus ustus, Aspergillus terreus, and
Chaetomium sp. The most dominant fungi found on leather
bookbinding were Penicillium oxalicum, P. rubrum, Penicil-
lium funiculosum, A. fumigates, A. niger, A. flavus, Aspergillus
versicolor, Aspergillus Wentii, and Fusarium sp.
Leather and paper are organic materials and are suscep-
tible to numerous biodeterioration processes, which
generally cause the loss of aesthetic properties and often
the irreversible degradation of important documents and
works of art [36]. The identified fungi are common on
historical papers and most of them are considered decom-
posed fungi for cellulosic and protein materials like paper
and leather [36]. Kowalik [37] and Held et al. [38] reported
that the bideterioration of cellulosic and proteineous mate-
rials by fungi depends mainly on the chemical composition
of the materials, its pH, its moisture content and the rela-
tive humidity of the environment, and the temperature
and the illumination. According to the promoting factors
mentioned above by Kowalik, Abdel-Maksoud [39] re-
ported that the conditions in most historical places in
Egypt are out of international standards that should be ap-
plied in museums, storehouses and libraries. It was found
by the measurement of pH (see below) of the manuscript
that the pH of paper and leather were acidic and fungi
prefer this level of pH. Some other factors encouraged
mold growth, such as a rapid fluctuation in relative
humidity associated with high changes in day and night
temperature.
4.3. pH measurement
It was noticed that the pH of leather was acidic, but the
acidity decreased more than the level in the normal state of
pH (3–5 pH). The pH of the leather bookbinding ranged be-
tween 2.5 and 2.9. The reduction of pH may be due to acid
accumulation in the leather, caused either by insufficient
removal of acidic residues from processing or by air-pollu-
tion, which can reduce the pH of the leather to fall below
3.0. Originally the effect of acid on leather is a general soft-
ening of the latter. After prolonged exposure, however, this
softening is transformed into a mellowness of the grain
and the leather structure become brittle. The next stage
is decay and total disintegration. The action of acids is
more destructive on vegetable-tanned leather [40].
The results of paper pH measurements showed that all
the paper studied showed acidic levels. The pH of studied
papers were 6.5, 5.6, 4.1 and 5.2 for original paper used
for writing, paper used in the restoration process, interior
lining paper and exterior lining paper, respectively. The re-
sults revealed that although the original paper was from
cotton fibers (as it will be explained later by SEM and
chemical analysis), the pH was acidic. This may be due to
some acids that formed within the papers, or those ab-
sorbed from the environment that were neutralized before
they had a chance to degrade the cellulose chains [41]. It
was stated that fibers made of cellulose chains degrade
when exposed to an acidic environment in the presence
of moisture. In this acid hydrolysis reaction, cellulose
chains are repeatedly split into smaller fragments so long
as the source of acid remains in paper. This acid hydrolysis
reaction produces more acid in the process, and the degra-
dation accelerates [41]. The results also revealed that the
interior lining paper had high acidity (4.1 pH). This may
be due to the internal factors such as the paper making
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process, and external factors such as environmental
conditions.
In the paper making process, especially in the late 19th
century, paper deterioration was further hastened by the
introduction of mechanically produced ground wood pulp.
These products, frequently not chemically purified, re-
sulted in weaker paper and the additional formation of
acids and peroxides that promote the aging process. The
paper made of ground wood pulp contained lignin, which
degrades to form acids and peroxides that further promote
the aging process. It can be added that the using of alum-
rosin sizing, especially in the mid 19th century onward,
causes the paper to eat itself from the inside out. In reac-
tion with the natural residual moisture in paper, alum
gradually breaks down to sulfuric acid, which attacks the
long chains of cellulose, breaking them into shorter and
shorter fragments. The paper steadily weakens until it fi-
nally becomes so brittle that it is unusable [41,42].
The external factors contain a series of chemical ele-
ments such as oxygen, nitrogen, ozone, and carbon dioxide.
These elements are responsible for the combustion,
fermentation, hydrolysis, and the oxidation of books and
documents. The atmosphere, especially of industrialized
zones, also contains a series of impurities, the results of
pollution or contamination, such as carbon dioxide, nitro-
gen dioxide, and sulfur dioxide. By-products of industrial
combustion which, catalyzed by metals, react with water
to form acids. The most important of these is sulfuric acid,
which leads to total weakness in the paper [42].
4.4. Investigation of the surface morphology by SEM and EDAX
analysis
Investigation of the paper by a SEM showed that fibers
of the original paper (Fig. 2A) seem to be from cotton.
Small amounts of filler materials appeared between the fi-
ber structures. Some contaminations (Fig. 2B) from stains
and dusts were noticed on the surface of the original paper.
Damages caused by insects (Fig. 2C) appeared in the form
of bores, and the tearing of paper fibers and deformation
Fig. 1. Aspects of deterioration found on Qur’an manuscript: (1A and 1C) aspects of deterioration of a bookbinding, (1B and 1D) aspects of deterioration of
papers.
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of the paper appearance was also noted. The restoration
paper (Fig. 2D) displayed advanced deterioration. Many
forms of deterioration such as dust, stains, and the random
distribution and destruction of the fibers were recorded.
For the exterior lining paper (Fig. 2E) dust and stains cov-
ered the surface of paper and the fiber structures is unrec-
ognizable. The amount of filler materials seems to be
greater than in the original paper. For the interior lining
paper (Fig. 2F) accumulated dust and large amounts of fil-ler materials were noted.
Investigation of leather bookbinding (Fig. 3) showed the
destruction and random distribution of the fiber struc-
tures, erosion of the fibers, and many bores. There was to-
tal deformation of the surface morphology.
The results obtained by EDAX analysis of stains found
on leather bookbinding supported the fact that the manu-
script was found under the ground of the destroyed mos-
que, and this was reflected by the elements present on
the surface of leather. Calcium was found in the high per-
centage of 27.77%. Magnesium, aluminum potassium, and
silicon were also found, and this may indicate that the
ground consisted of lime with traces of clay minerals and
sand. Sulfur may act as a contaminant from the surround-
ing environment or from the manufacturing process of
leather bookbinding. Sodium also was found and may have
derived from the burial environment of the manuscript.
The sodium was further identified as sodium chlorideand it was found on all of the paper pages of the manu-
script. This type of salt led to the erosion of the surface
of both the papers and leather. The source of sodium chlo-
ride in the case study may be from saline in the soil and
groundwater, air pollution, and human contaminants. Salt
damage takes place when evaporation takes place, leaving
the salt to grow as crystals within the pores of the leather
or paper. The growth pressure of developing crystals is
very high and sufficient to cause erosion on the leather
Fig. 2. Investigation of deteriorated of paper by SEM: (A) original paper manuscript; (B) original paper with some stains and dusts; (C) original paper with
damaged caused by insects; (D) restoration paper; (E) exterior lining paper; (F) interior lining paper.
Fig. 3. Investigation of the surface morphology of leather bookbinding by SEM and analysis of stain found on the leather surface by EDAX.
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or paper sheets. The final result is total deformation of the
surface.
4.5. X-ray diffraction and EDAX analysis of ink and pigments
used
4.5.1. Gold color
It was clear from X-ray diffraction (Fig. 4A) and EDAX(Table 1) that the gold color was from gold leaf or gold
shell. Gold leaf was applied before any painting was done
because it would stick to the medium used to suspend
the paints. Shell gold was also used to illuminate manu-
scripts, it is made of powdered gold suspended with
gum. It was cheaper than gold leaf and could be applied
with a pen or brush. Shell gold allowed for finer detail
and could be applied after the paint.
4.5.2. Red color
The data obtained (Fig. 4B and Table 1) showed that the
red color, Vermillion, was formed from red mercuric sul-
fide obtained from cinnabar, the principle ore of mercury,or artificially by heating sulfur and mercury together [43].
4.5.3. Blue color
It was clear from Fig. 4C and Table 1 that the blue color
consisted mainly of cobalt oxide with aluminum oxide. Co-
balt blue was useful in all techniques, as well as being
lightproof. It needed a binder like gum Arabic [43]. Sodium
chloride was also identified, and was found with paper
support of the manuscript.
4.5.4. Black ink
Iron gall ink is the most important ink used in old
manuscripts. Many recipes exist for manufacturing black
ink (Fig. 4D), and usually contained a mixture of carbonand iron gall. Therefore a wide range of different compo-
nents and impurities exist for historical inks. The results
from the analysis of black ink showed the use of iron and
sulfur (iron sulfate), as seen in Table 1. In this study the
ink used may be a mixture of carbon and iron gall ink.
The significant amount of calcium, potassium, and magne-
sium in the black ink samples demonstrates the possible
use of gum Arabic as a binding media [13].
4.6. Identification of pigment binder by FTIR
The binder used with the black ink and pigments (Fig. 5)
was identified as gum Arabic after a comparison with thecontrol sample of pure gum Arabic. Bands at 2965–
2880 cm1 and 1415–1380 cm1 are assigned to CH3 and
CH2, which by its chemical composition is similar to natu-
ral hydrocarbons such as gum Arabic. A very strong band at
1030 cm1, due to C–O, indicated the characteristics of
polysaccharides. The identified gum Arabic also contained
Fig. 4. X-ray diffraction pattern of ink and pigments used on the paper of the manuscript.
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moderately strong bands at 1625–1630 cm1. Derrick et al.
[44] explained that bands at 1625–1630 cm1 is partially
associated with intramolecularly bound water and par-
tially due to the presence of a carboxyl group.
4.7. Measurement of hydrothermal stability of leather
bookbinding
The hydrothermal stability of collagen fibers, shrinkage
by heating in water, is a particularly good measure of the
strength or quality of leather and skin materials and the
degree of their deterioration [7]. Raw collagen, when
heated in water, shrinks at about 65 C [45]. Chemical
cross-links introduced into the collagen by tanning agents
raise the shrinking temperature depending on the type of
tanning material and the nature of the process employed
[46]. The shrinkage temperature of vegetable tanned
leather is between 75 and 85 C [46]. Larsen et al. [17] re-
ported that the phrase ‘shrinkage activity’ is used to denote
any observable shrinkage process going on in a fiber, thin
or thick, and the phrase ‘intensity of the shrinkage activity’
as a qualitative measure of the number of the observable
shrinkage processes taking place at a small temperature
interval.
The results (Fig. 6) revealed that the average five mea-
surements of the shrinkage temperature of the historical
leather bookbinding sample was 64 C. The shrinkage tem-
perature of the historical leather decreased from 15 C to
25 C compared with the control samples. Haines [47] ex-
plained that the reason for the shrinkage temperature is
that the backbone chains of the molecule exist in an ex-
tended form, held in this form by hydrogen bonding. Whencollagen is heated there is a point where the energy input
exceeds that of the hydrogen bonding. Then there is a sud-
den release from the extended form and the fibers shrink
to a rubber-like consistency. Only the remaining covalent
and salt bonds hold the collagen molecules together and
prevent the shrunken collagen from immediately going
into solution.
The decreased shrinkage temperature in the historical
leather bookbinding may be due to ageing conditions.
Haines [46] reported that ageing conditions that bring
about hydrolytic or oxidative degradation of the collagen
cause breaks in the backbone chain of the molecule and
changes to the chemical composition of the side chains.These both lead to a reduction in the shrinkage
temperature.
Chahine [20] said that there are different factors that
influence the shrinkage temperature. They can be classified
into biological and non-biological factors. Biological factors
relate to the nature of animal species, its living conditions,
environment, age of the animal, and from where on the
body the sample was taken. Furthermore, it was shown
by different authors that the shrinkage temperature (Ts)
is related to the amino acid content of collagen, their posi-
tion in the chain, and their hydroxylation. Fish skin, for
example, has a lower content of amino acids and a lower
Ts than mammal skin. Non-biological factors include thenature of the heating medium, ionic environment, salts,
tanning, and ageing.
4.8. Identification of animal skin used for the bookbinding
It was clear by a study of the grain surface of the book-
binding by SEM and the comparison with the study of
Haines [32] (Fig. 7A and B) that the type of skin used for
the bookbinding was goat skin. The leather surface was
smoothed and glazed. The coarse follicles were in the form
of groups. There was a wide and smooth surface between
these groups. The grouping of coarse and fine follicles
was easily recognized.
Table 1
EDAX analysis of ink and colorants used on paper manuscript.
Color Elements (wt%) Total
Na Mg Al Si P S Cl K Ca Ti Fe Cu Zn Hg Au O Co
Gold 1.37 0.22 0.43 0.58 4.15 0.27 0.15 0.60 0.88 – 0.28 1.32 1.24 – 71.17 17.33 – 100
Blue 9.20 – 9.79 18.93 0.71 4.43 1.54 2.12 7.66 – 0.78 – – – – 42.63 2.22 100
Red 0.80 3.03 3.65 10.67 – 6.02 3.15 2.05 19.37 – 0.73 0.78 0.30 28.50 – 20.95 – 100
Black 2.65 2.45 3.24 11.32 1.19 4.59 9.35 4.09 18.40 0.33 3.82 0.82 0.62 – 37.12 100
Fig. 5. FTIR analysis of gum Arabic binder used with ink and pigments:
(1) control, (2) gold color, (3) red color, (4) black ink, and (5) blue color.
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4.9. Identification of tanning material by High Performance
Liquid Chromatography (HPLC)
The identification of the tannin type used with the book-
binding was determined by the comparison of the retention
times and UV spectra between Acacia Arabica powder and
tannin material extracted from leather bookbinding. It
was clear from the data obtained (Fig. 8A and B) that there
was a close similarity in the retention times for the effective
peaks of tanning extracted from the bookbinding studied
and the tanning extracted from Acacia Arabica powder.
The close similarity in the retention times ranged from be-
tween 1.727 and 4.454 min. The results proved that there
was a similarity in the spectral characterization of individ-
ual tanning peaks. The retention times (min) of the effective
peaks of Acacia Arabica powder (Fig. 8A) were 1.918, 2.268,
2.623, 3.820 and 4.454, respectively, and the UV spectra of
these peaks were 235.20 nm, 257.53 nm, 292.15 nm,
369.85 nm and 456.20 nm, respectively. The retention
times (min) of the effective peaks of the extracted tannin
from the leather bookbinding (Fig. 8B) were 1.727, 2.270,
2.670, 3.804 and 4.330 min, respectively, and the UV spec-
tra of these peaks were 230.10 nm, 260.45 nm, 296.65 nm,
365.28 nm and 450.20 nm, respectively. It should be noted
that most of the intensity of the tanning material extracted
from Acacia Arabica powder was higher than tanning mate-
rial extracted from vegetable-tanned leather bookbinding.
The clarity and high intensity of peaks of thespectra for Aca-
cia Arabica powder compared to those for the vegetable-
tanned leather bookbinding may have been due to the
mechanism of deterioration, oxidation or hydrolysis pro-
cesses, of the vegetable-tanned leather bookbinding. This
comparison clearly shows that the tanning material used
on the bookbinding has some similarity to Acacia Arabica.
Fig. 6. Microscopic pictures of vegetable tanned leather bookbinding: (A) shrinkage temperature of the sample at room temperature at 22 C, beforeshrinkage; (B) shrinkage activity of individual fibers at 48 C; (C) shrinkage activity in more than one fiber at 55 C; (D) shrinkage activity in some fibers at
59 C; (E) the final shrinkage activity at 64 C; (F) shrinkage temperature measurement of different sample of the leather bookbinding.
Fig. 7. Identification of animal skin: (A) goat skin (after Haines, 1981), (B) goat skin from the historical bookbinding.
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4.10. Chemical analysis of paper
The results of the chemical analyses for ash, lignin anda-cellulose are presented in Table 2. A high percentage
was recorded for a-cellulose in the original paper (57%)
and a low percentage was obtained from interior lining
paper (25%). For lignin and ash, a low percentage was
recorded with the original paper and the high percentage
was obtained from the interior lining paper. The paper
used in the restoration process contained higher percent-
age of lignin and ash than the original paper, but these per-
centages were lower than the interior and exterior papers.
These results argue that pure cellulose was used in the
manufacturing of the original paper and wood fibers may
have been used in the other papers. Due to the high cellu-
lose content and low lignin and ash contents of the originalpaper, it was relatively more resistant to the surrounding
environmental conditions than the other papers. Ververis
et al. [47] reported that paper made of wood fiber pulp is
weak and its resistance towards the surrounding environ-
mental conditions is poor.
5. Conclusion
This study proved that the manuscript studied suffers
from deterioration caused by surrounding environmental
conditions. By visual assessment and by investigation the
surface morphology (SEM), many aspects of deterioration
were noted on the surface of the papers or leather book-
binding, such as crystallization of the sodium chloride,
holes caused by insects, wrapping, erosion of tanning
material, and missing parts. The surrounding environmen-
tal conditions with the materials used in the manufactur-
ing of this manuscript play an important role in the
growth of fungi. The most dominant fungi were Aspergillus
sp., Penicillium sp., Chaetomium sp., and Fusarium sp. The pH
value of the paper and leather bookbinding were lower
than their values in the normal condition. This may due
to the manufacturing process and the surrounding
environmental conditions. The shrinkage temperature of
leather bookbinding was reduced compared to normal
the values of the shrinkage temperature of vegetable-
tanned leather. The reduction of the pH value of the leather
bookbinding below 3 pH may play an important role in the
reduction of the shrinkage temperature. The shrinkage
temperature of leather, gave a clear indication that the
manuscript suffers from deterioration. It proved that the
deterioration mechanism by hydrolysis or oxidation pro-
cesses caused breaks in the backbone chain of the molecule
and changed the chemical composition of the side chains.
The black ink was a mixture of carbon and iron gall. The
gold color was from gold leaf or gold shell, the red color
was from mercuric sulfide, and the blue color was from co-
balt oxide. The binder used with black ink and pigments
was gum Arabic. The scanning electron microscope inves-
tigation and chemical analysis of paper contents proved
that the original paper used as a support of writing mate-
rials was from cotton, but the wooden fibers may be used
in the exterior and interior papers used with the leather
bookbinding. Goat skin was the animal skin used for the
leather, and Acacia Arabica was the tanning material used
with the bookbinding studied.
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Fig. 8. HPLC elution profile: (A) New tanning material extracted from Acacia Arabica. (B) Tanning material extracted from vegetable-tanned bookbinding.
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