Journal of Cultural Heritage 12 (2011) 149–156

Original article

Tannins characterisation in new and historic vegetable tanned leathers fibres by

spot tests
Lina Falcão , Maria Eduarda M. Araújo ∗
Chemistry and Biochemistry Centre, Faculty of Sciences, University of Lisbon, Campo Grande, Edifício C8, 1749-016 Lisboa, Portugal

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

Article history: This paper describes the adaptation and evaluation of three chemical tests for tannins characterisation in
Received 27 May 2010 vegetable tanned leathers. Tests were performed on fibres of new leathers tanned with different known
Accepted 12 October 2010 vegetable tannins and historic leathers. Rhodanine test, nitrous acid test and acid butanol test, developed
Available online 14 December 2010
to identify, respectively, gallotannins, ellagitannins and condensed tannins, are described. Ferric test and
vanillin test, the two traditional tests used for vegetable tanned leathers characterisation, were also per-
Keywords: formed and their usefulness discussed. Gallic acid, ellagic acid and catechin, structural constituents of the
Leather
different classes of tannins were also tested. Results of the developed methodology allowed the identi-
Vegetable tanning
Tannins
fication of tannins’ chemical class in new and historic leather samples studied. Data obtained permitted
Colorimetric tests to verify the information on tanning materials used in new leathers. Vegetable tanning technology was
Restoration leathers confirmed in historic samples and tannins were characterised. This study shows that these tests are use-
Historic leathers ful and can be a valuable source of information to evaluate new vegetable tanned leathers quality for
Key-guide conservation and restoration purposes as well as historic leathers tanning technology.
© 2010 Elsevier Masson SAS. All rights reserved.

1. Research aims Tannins, organic compounds present in barks, leaves, wood or

The aim of this work is to present a new and straightforward
methodology developed to identify the type/class of tannins in new
and historic vegetable tanned leathers. This information is valu-
able to comprehend leather technology, degradation susceptibility
and condition. Methodology involved the adaptation of chemical
and colorimetric tests, formerly used for the analysis of tannins in
vegetable materials, to be used principally in conservation studios
or laboratories, as a routine quality evaluation of commercial veg-
etable tanned leathers for conservation and restoration purposes
as well as in technology and conservation assessment of historic
leathers.
2. Introduction
Leather production is an ancient technology that has been
described as man’s first manufacture process [1]. It comprises the
transformation of raw putrescible animal skin (and hide) into a use-
ful material, which is less permeable to water, more resistant to
abrasion and has enhanced stability to microbial attack and heat.
This is due to the ability of some organic and inorganic compounds
to react with the main skin constituent: the collagen protein [2].
∗ Corresponding author. Tel.: +351 217500968/+351 217500188;
fax: +351 217500968/+351 217500188.
E-mail address: eduaraujo@fc.ul.pt (M.E.M. Araújo).
galls of many plants, have been employed – either alone or in com-
bination with other materials – since Neolithic time to retard decay
and to convert skin to leather [1,3,4].
This technology, developed in the Mediterranean region [5]
and known as vegetable tanning, was during centuries the dom-
inant leather making process in Europe until it has gradually been
replaced by chromium mineral tanning by the end of the 19th cen-
tury [1,6–8]. And although the term tanning was often reserved for
leather production with tannins since the development of other
processes in the 19th century, the word has been applied for all
different processes of leather making [1].
Vegetable tanned leather has been widely used for the pro-
duction of footwear, saddlery, harness, bookbinding, upholstery or
wall-hangings [4,9,10] and it is the most common type of leather in
western cultural heritage [1,11]. Frequently, these leather objects
present degradation problems and require conservation. Treat-
ments such loss compensations, reinforcements or, in more critical
cases, partial or complete replacement of heavily degraded veg-
etable tanned leather often requires the application of a similar
type of leather [12–14].
Tannins are a complex and heterogeneous group of polypheno-
lic secondary metabolites of higher plants with molecular weight
between 500 and 20,000 Da. They share the ability to bind to and
precipitate proteins, alkaloids and polysaccharides. This particular
reactivity with proteins is named astringency [15,16].
Tannins composition and concentration vary considerably with
the specie, age and part of the plant. Depending on the source

1296-2074/$ – see front matter © 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.culher.2010.10.005
150 L. Falcão, M.E.M. Araújo / Journal of Cultural Heritage 12 (2011) 149–156
used for tanning, extracted tannins confer different organoleptic
and chemical properties to the produced leathers [17–19]. Past
observations and recent experimental studies clearly showed that
the chemical characteristics of tannins play an important role on
leather stability [20].
Moreover, to be efficient in leather making tannins molecu-
lar weight must be between 500 and 3000 Da. Higher molecular
weights are unable to penetrate into the skin’s fibre structure and
tend to be water insoluble [1,2].
The main vegetable sources used for tanning [17,18,21,22] are
presented in Table 1.
Depending on their chemical composition, tannins can be clas-
sified in two main groups: condensed (or proanthocyanidins) and
hydrolysable tannins [5,15]. Plants, which have a strong capacity to
biosynthesise condensed tannins, rarely metabolise considerable
amounts of hydrolysable tannins, and vice-versa [23]. Bark from
Quercus petraea is one exception example, since accumulates both
types of tannins [24].
This traditional classification, defined by Freudenberg in 1920
[15,23], has been enlarged to two more classes in the last decades:
complex tannins and phlorotannins. The last one is a small special
group isolated from marine brown algae with no importance for
leather making [5].
Condensed tannins are oligomeric or polymeric flavonoids con-
sisting of flavan-3-ol (catechin) units, which are commonly linked
through C4→C8 positions, but C4→C6 links may also occur (Fig. 1).
Upon treatment with hot alcoholic acid solutions, this class of tan-
nins yields red-coloured anthocyanidins and insoluble polymeric
phlobaphenes. Examples of important condensed tannins for veg-
etable tanning are extracted from quebracho wood and mimosa
bark [2,22].
Condensed tannins produce reddish-brown leathers, which
tend to darken when exposed to light [19,25,26]. These leathers
are also prone to adsorb atmospheric pollutants, among which sul-
phur dioxide (SO2 ) is the most deleterious, promoting collagen acid
hydrolysis [1,20].
Hydrolysable tannins consist of a polyol core, being d-glucose
the commonest, multi-esterified with gallic acid (3,4,5-trihydroxyl
benzoic acid) or derivatives. These tannins undergo on hydrolytic
cleavage into their components, hence the name. Depending on
the phenolic acids formed by hydrolysis they are subclassified in
gallotannins (Fig. 2) and ellagitannins (Fig. 3).
Gallotannins are the less common in nature and when degraded
by hydrolysis yield gallic acid and the polyol core. They are charac-
terised by glucose core esterified with 10–12 gallic acid units, either
directly or by depside bonds between gallic acid units (Fig. 2b).
Those extracted from sumac leaves or tara pods are important
examples in vegetable tanning [2,22].
Ellagitannins are characterised by glucose core esterified with
at least one unit of hexahydroxydiphenic acid, which is formed
through oxidative coupling between two gallic acid units. Upon
hydrolysis, hexahydroxydiphenic acid is released, and sponta-
neously lactonizes forming ellagic acid (Fig. 3a and b). Most
ellagitannins are mixed esters: glucose hydroxyls not linked to hex-
ahydroxydiphenic acid are esterified with gallic acid and, in some
cases, with chebulic acid. Extracts of chestnut wood are the most
common ellagitannins used for vegetable tanning [2,22], but the
use of oak wood has been also historically important [21,27,28].
Hydrolysable tannins produce pale yellow, greenish or brown
leathers, which are typically lightfast and considered more sta-
ble than those produced with condensed tannins [19,20,25,26].
For conservation and restoration purposes, only vegetable tanned
leathers produced with hydrolysable tannins are recommended
[29,30].
Complex tannins comprise flavan-3-ol units glycosidically
bound to gallotannins or ellagitannins units and are minor con-
stituents of vegetable sources used in leather production [2,15].
They have been detected in Q. petraea bark [24] and in chestnut
commercial tanning extracts [31].
The analysis of tannins is a relatively common procedure in
leather conservation. Tannins characterisations have been impor-
tant not only to elucidate aspects on historic leathers composition,
technology [25,32–36], and condition [34], but also to evaluate the
suitability of new vegetable tanned leathers for conservation pur-
poses [29,30,33].
Tannins characterisation in leathers is currently based on
two spot tests: ferric test [32,37] and vanillin test [29,33,35].
These chemical tests are employed for a fast preliminary mate-
rial characterisation [37,38], especially by leather conservators.
More accurate analysis is based on the instrumental technique of
high performance liquid chromatography (HPLC), which is mainly
used for interdisciplinary studies in the conservation science
field [20,30].
Ferric test has been used to detect if tannins are (or not) present
and vanillin test to detect condensed tannins in leather fibres.
Considering the diversity of vegetable sources and the enlarged
classification of tannins used for leather production, the informa-
tion obtained with both chemical tests is very limited.
Chemical tests, such as rhodanine, nitrous acid and acid butanol,
have been used for tannins analysis in plant tissues [16,39,40].
Adaptation of those tests for leather fibres analysis, as well as their
utilization as spot tests, was performed and herein described. This
paper presents the assessment of the above three chemical tests for
tannins characterisation in fibres collected from new and historic
leathers.
3. Experimental
Qualitative characterisation of tannins was performed on
leather fibres samples by semi-micro analysis with five chemical
and colorimetric tests: ferric, vanillin, rhodanine, nitrous acid and
acid butanol. The first three tests were carried out in a spot test
porcelain plate and the former two in small-capped glass vials.
Analysed fibres, approximately 40 mg, were collected in the
reticular layer (corium) of leathers samples.
Ferric test and vanillin test procedures were used as described
by Larsen et al. [29]. Rhodanine, nitrous acid and acid butanol tests
were adapted from Hagerman [39]. Results were evaluated visually
and compared with controls (blank tests with the same amount
of leather fibres, or standards, were performed with all reagents
except the one responsible to the formation of the coloured prod-
uct).
3.1. Samples
Nine vegetable tanned leathers were analysed. Six leathers were
recently produced with tannins extracted from diverse known
plant sources, which comprise the different chemical classes of tan-
nins. The other three leathers were historic and dated from the 19th
century: one bookbinding leather (HL1) and two different kinds of
upholstery leathers: a plain one (HL2) and the other a morocco type
(HL3).
New vegetable tanned leathers (NL1-6) were obtained in British
tanning industries or supplied by German institutions (Table 2).
Information on the tanning materials, provided with the samples,
distinguished two kinds of vegetable tanned leathers: four leather
samples produced with one single vegetable tanning material,
specifically, mimosa (NL1), valonia (NL2), oak bark from Q. petraea
(NL3) and chestnut (NL6), and the other two samples with mixtures
of tannins. NL4 was tanned with sumac after being imported from
Nigeria where it had been tanned with bagaruwa–local term for
 L. Falcão, M.E.M. Araújo / Journal of Cultural Heritage 12 (2011) 149–156 151

Table 1
Main sources of vegetable tannins used for leather production.

Family Specie(s) scientific name Part of the plant used Tanning material common name Chemical classification

Anacardiaceae Schinopsis lorentzii, Schinopsis balansae Heartwood Quebracho Condensed tannins [22,31]
Rhus typhina, Rhus coriaria Leaves Sumac Gallotannins [22,31]
Rhus semialata (syna R. chinensis) Galls Chinese Gallotannins [22,31]

Combretaceae Terminalia chebula Fruits Myrabolans Ellagitannins [22]

a
Fagaceae Castanea sativa (syn C. vesca, C. Wood Chestnut Ellagitannins [22,31]
vulgaris)
Quercus aegilops (syna Q. ithaburensis Acorn cups Valonia Ellagitannins [22,31]
subsp. macrolepis)
Quercus infectoria Galls Turkish, aleppo Gallotannins [22,31]
Quercus petraea (syna Q. sessiliflora) Wood Oakb /sessile oak wood Ellagitannins [28]
Bark Oakb /sessile oak bark Condensed tannins and ellagitannins [24]
Quercus robur (syna Q. pedunculata) Wood Oakb /pendunculate oak/English oak Ellagitannins [28]
wood
Bark Oakb /pendunculate oak/English oak Condensed tannins [45]
bark
Leguminosae Acacia mearnsii (syna A. mollissima Bark Mimosa Condensed tannins [22,31]
auct.)
Caesalpinia coriaria (syna Poinciana Pods Divi-divi Ellagitannins [22,31]
coriaria)
Caesalpinia spinosa (syna C. pectinata, C. Pods Tara Gallotannins [22]
tinctoria)
a
Taxonomy synonyms in, USDA, ARS, National Genetic Resources Program, Germplasm Resources Information Network (GRIN) [Online Database]. National Germplasm
Resources Laboratory, Beltsville, Maryland. URL: http://www.ars-grin.gov/cgi-bin/npgs/html/taxgenform.pl?language=en (05 July 2010)
b
Oak, common name frequently used to many Quercus species.

Acacia nilotica pods [41]. NL5 was tanned with sumac and retanned
with a mixture of chestnut and quebracho.
3.2. Reagents and solvents
The reagents used were ferric chloride (Panreac), vanillin (4-
hydroxy-3-methoxybenzaldehyde), sodium nitrite (Fluka), potas-
sium hydroxide (Fluka), rhodanine (2-thio-4-ketothiazolidine, Alfa
Aesar), ferric ammonium sulphate (Merck) and 37% hydrochlo-
ric acid (Panreac). Reference compounds, gallic acid, ellagic acid,
tannic acid and catechin, were supplied by Sigma. All solvents
used, n-butanol, ethanol, methanol and pyridine, were of analytical
grade.
Ferric reagent was an aqueous solution of 2% (w/v) ferric
chloride (FeCl3 ). Vanillin reagent contained 1% (w/v) vanillin
in ethanol. Acid butanol reagent was prepared mixing 95 mL

Fig. 1. General structure of condensed tannins: (R=H or OH): a) a flavan-3-ol monomer, catechin (R=H).
152 L. Falcão, M.E.M. Araújo / Journal of Cultural Heritage 12 (2011) 149–156

Table 2
Results of colorimetric tests.

Samples Description Ferric test Vanillin test Acid butanol test Nitrous acid test Rhodanine test

New
NL1 Industrial tanning in UK with mimosa +Dark grey + Red + Crimson − Orange − Orange
NL2 Laboratorial tanning in Germany with valonia +Dark grey − No colour − Yellow + Blue − Orange
NL3 Laboratorial tanning in Germany with Quercus patraea bark +Dark grey + Red + Crimson + Blue − Orange
NL4 Rural tanning with bagaruwa and industrial tanning in UK with sumac +Dark grey + Red + Red orange − Orange + Pink
NL5 Laboratorial tanning in UK with sumac, chestnut and quebracho +Dark grey +Brownish red + Red orange + Blue +Brownish pink
NL6 Industrial tanning in UK +Dark grey + Red + Crimson + Blue +Brownish pink

Historic
HL1 19th century bookbinding, vegetable tanning +Dark grey − Yellow + Red orange −Reddish brown + Pink
HL2 19th century plain upholstery leather, vegetable tanning +Dark grey − Light brown + Crimson − Brown − Orange
HL3 19th century morocco upholstery leather, vegetable tanning +Dark grey − No colour − Yellow −Reddish orange + Pink

+: positive test; −: negative test.

of n-butanol with 5 mL of 37% HCl. Iron reagent contained 2%
(w/v) ferric ammonium sulphate (FeNH4 (SO4 )2 ·12H2 O) in 2N
HCl. Rhodanine reagent contained 0.667% (w/v) rhodanine in
methanol.
3.3. Ferric test for tannins detection
3.4. Vanillin test for condensed tannins detection
A sample with approximately 1 mg of leather fibres was used.
Two drops of vanillin reagent were added to the sample. When
completely absorbed, two drops of 37% HCl were added. The
development of a red colour indicates the presence of condensed
tannins.

A sample with approximately 1 mg of leather fibres was tested.
Sample was first hydrated with water. Water excess was removed
with blotting or filter paper. Then, two drops of ferric reagent were
added to the sample. A grey or black colour formation indicates the
presence of tannins.
3.5. Acid butanol test for condensed tannins detection
In a small glass vial 2.4 mL of acid butanol reagent was added to
5 mg of leather fibres. Then, 40 ␮L of iron reagent was added and the
mixture was heated up to 100 ◦ C for 50 min. A progressive forma-

Fig. 2. General structure of gallotannins: a): gallic acid structure; b): digallic acid structure (m-depside bond).
 L. Falcão, M.E.M. Araújo / Journal of Cultural Heritage 12 (2011) 149–156 153

Fig. 3. Structure of casuarictin, an example of an ellagitannin: a): hexahydroxydiphenic acid structure; b): ellagic acid structure.

tion of a red-orange to red-crimson product indicates the presence 3.8. Standards testing
of condensed tannins.
Compounds such as gallic acid, ellagic acid and catechin, struc-
tural components of the different types of tannins, were also tested
3.6. Nitrous acid test for ellagitannins detection
for comparative purposes. Tannic acid, a gallotannin, was also
tested.
In a small glass vial 2 mL of pyridine was added to 5 mg of leather
The standards were prepared as follows: aqueous solutions of
fibres. Then 150 ␮L of 37% HCl was added and the mixture heated
0.1 mg/mL gallic acid and tannic acid, an aqueous suspension of
up to 30 ◦ C. After 5 min 150 ␮L of 1% (w/v) sodium nitrite (NaNO2 )
0.5 mg/mL ellagic acid and a solution of 0.1 mg/mL catechin in
in water was added, mixed and incubated for 20 min at 30 ◦ C. A
methanol.
progressive blue colour formation indicates the presence of ellagi-
Two drops of each compound were tested using the procedures
tannins.
described for leather fibres. Results are presented in Table 3.

3.7. Rhodanine test for gallic acid and gallotannins detection

A sample with approximately 1 mg of leather fibres was tested.
Two drops of rhodanine reagent were added to the sample. Five
minutes later, two drops of aqueous 0.5N potassium hydroxide
(KOH) were added. A progressive pink colour formation within
5 min indicates the presence of gallic acid.
4. Results and discussion
4.1. Ferric test for tannins detection
Ferric test is specific for phenolic compounds and not only
tannins. It is based on the principle that phenolics react with
154 L. Falcão, M.E.M. Araújo / Journal of Cultural Heritage 12 (2011) 149–156
Table 3
Results of standards with colorimetric tests.
Standards Ferric Vanillin Acid butanol Nitrous acid
Gallic acid Black – – – Pink
Ellagic acid Black – – Red
Catechin Black Red – Yellow
Tannic acid Black – – – –
– No colour formed.
iron salts forming a blue or green black (or grey) product
[17,25].
All tested samples showed a positive result for phenolics
(Table 2). Nevertheless, phenolic compounds found in leather can
either be due to tannins or some natural dyes [33]. New vegetable
tanned leathers were not dyed. Therefore, it can be safely concluded
that the phenolics detected were tannins.
Visually it is not always clear if historic leathers have been
dyed or not, because the colours may have faded. Analysed fibres
of historic samples were yellowish and brown, colours associated
to vegetable tanning, but it is not known if they have been dyed.
Therefore the positive ferric test by itself just gives an indication of
vegetable tannins presence, which confirmation can only be sup-
ported by the other performed tests.
Some literature refers ferric (and ferrous) test can be used to dis-
tinguish condensed from hydrolysable tannins [17,25]. Condensed
tannins form green black products and hydrolysable form blue
black products. This subtle distinction is very difficult to observe
in leather fibres, specially, if mixtures of different types of tannins
are present and diverse coloured fibres are being compared. No
clear distinction was visually perceived in the analysed samples.
4.2. Vanillin and acid butanol tests for condensed tannins
detection
Vanillin test has been widely used to detect condensed tan-
nins but the reaction is not specific for those phenolic compounds
[39,42]. In acid conditions, vanillin reacts specifically with meta-
substitued flavanols to form a red product. Widely distributed
flavanols such as catechin (Table 3) and epicatechin, condensed tan-
nins monomers, also react with the disadvantage they give higher
colour yield than the condensed tannins. Furthermore, it is very
sensitive to the presence of water that quenchs colour yield. In addi-
tion the 5-deoxy proanthocianidins, like quebracho, do not produce
an intense colour with this reagent. Reactivity with vanillin is not
considered sufficient evidence for the presence of condensed tan-
nins [39]. Moreover it has been reported [33] that vanillin reacts
reluctantly with aged leathers, becoming very difficult to clearly
identify the presence of condensed tannins.
Acid butanol test is specific for condensed tannins. It promotes
an oxidative depolymerisation of condensed tannins in heated
acidic alcohol to yield correspondent red anthocyanidins. The
red colour can vary significantly depending on the anthocyanidin
formed [16] and on the position of interflavan links (C4→C6 were
found to be more resistant to cleavage than C4→C8) [39,42].
Vanillin and acid butanol results were coherent for new leathers
(Table 2). All new samples, with exception of NL2, gave positive
results.
NL6 positive identification was not expected since it was sup-
plied as having being tanned with chestnut tannins, a source of
ellagitannins (Table 1). Considering both vanillin and acid butanol
tests were unequivocally positive, the presence of condensed tan-
nins cannot be considered an interference of the experimental
procedure. This sample must have been tanned with a vegetable
source rich in condensed tannins.
The three historic leathers tested negative with vanillin test, but
HL1 and HL2 showed a positive reaction towards acid butanol test,
Rhodanine which is not surprising bearing in mind that vanillin test gives less
significant results with aged samples [33]. Although acid butanol
test requires more time to perform it is a good alternative to the
Yellow
vanillin test [39].
–
4.3. Nitrous acid test for ellagitannins detection
Ellagitannins react with nitrous acid, obtained by dissolving
sodium nitrite in a diluted acid, forming a red or pink colour, which
slowly changes to purple or blue [40,43].
As expected, NL2, NL3, NL5 and NL6 tested positive (Table 1 and
Table 2).
All historic samples tested negative. Although a formation of
a reddish/brown colour has developed no blue final product was
observed (Table 2).
Nitrous acid test can also be used to detect ellagic acid [43], an
ellagitannins degradation product. With this reagent ellagic acid
forms a red product while other phenolics can form yellow or
orange products (Table 3). Ellagic acid can be present in aged leather
due to acid hydrolysis promoted by moisture and acidic pollutants.
The red colour formed in tested historic samples indicates ellagic
acid may be present.
4.4. Rhodanine test for gallic acid and gallotannins detection
Rhodanine test, developed by Inoue and Hagerman for gallotan-
nins determination, it is based in the determination of gallic acid
released by hydrolysis of tannins [44]. In basic solution, rhodanine
reacts with the vicinal hydroxyl groups of free gallic acid producing
a red complex. Ellagic acid, catechin and tannic acid do not interfere
with this test (Table 3).
This test has been previously intended to detect directly gal-
lotannins in leathers fibres [33]. Samples were visually compared
before and after hydrolysis with the aim to distinguish different
amounts of gallic acid. But, when the test was performed, no visual
difference was observed. Within this work, rhodanine test was
adapted to detect free gallic acid in fibres and, indirectly, to estimate
gallotannins, without hydrolysing samples, by comparing results
with acid butanol and nitrous acid tests (Fig. 4). The presence of
free gallic acid in leathers may be due not only to gallotannins, but
also to ellagitannins [44], complex tannins and even condensed tan-
nins degradation [42]. Therefore, an unequivocal assumption of a
leather tanning with gallotannins is only possible if ellagitannins
and condensed tannins are not detected (Fig. 4).
NL4, NL5 and NL6 tested positive for gallic acid (Table 2), which
agrees with the tanning products used (Table 1). HL1 and HL3 also
tested positive.
4.5. Overall evaluation of spot tests on tested samples
Regarding new leathers, NL1-5 results are in agreement with
the information of the tanning agent provided by the supplier.
However, results of NL6 indicate that information provided about
tanning agent was incomplete or even incorrect since condensed
tannins were detected (all tests gave positive results meaning that a
mixture of tannins is present, Fig. 4). Indeed, if only chestnut (ellag-
itannins source) had been used, condensed tannins could not be
present in NL6.
Notwithstanding the small number of historic leathers samples
available in this study, one can say that tests used can also be use-
ful to analyse aged and historic materials, using a small amount
of leather fibres. Regarding HL1 sample results (tested positive for
condensed tannins and gallic acid, and negative for ellagitannins)
indicated that this leather was produced with a source of condensed
 L. Falcão, M.E.M. Araújo / Journal of Cultural Heritage 12 (2011) 149–156 155

SAMPLE TESTS RESULTS INTERPRETATION

All negative No tannins are present

All positive Mixture of tannins

BT negative
NT positive Ellagitannins or a mixture
ACID BUTANOL RT positive of gallo- and ellagitannins

TEST (BT) BT negative

NT negative Only gallotannins
+ RT positive (eg. sumac, tara)
FIBRES
NITROUS ACID
BT negative Only ellagitannins
FROM NT positive (eg. valonia, chestnut)
TEST (NT)
RT negative
THE CORIUM
+
BT positive Condensed tannins
RHODANINE NT negative or a mixture of gallotannins
RT positive and condensed tannins
TEST (RT)
BT positive
NT positive Mixture of condensed
RT negative tannins and ellagitannins

BT positive
NT negative Only condensed tannins
RT negative (eg. mimosa)

Fig. 4. Key-guide for the characterisation of the tannin type in leather fibres.

tannins that include gallic acid, and became free with deteriora-
tion [42] or with a mixture of condensed tannins and gallotannins
(Fig. 4). Regarding HL2 sample (only tested positive for condensed
tannins), this information contributed to a better understanding
of sample tanning production since it was collected from a German
chair whose leather was probably tanned with oak bark (Dr. Schulze
personal information). Considering that oak bark from Q. robur was
very used in Central Europe and is a source of condensed tannins,
this hypothesis was not discarded. In HL3 sample, a distinctive
morocco type of leather whose tanning with sumac or galls (both
gallotannins) has been historically described [21], only gallic acid
was detected, which supports historic information.
Carrying out spot tests with chemical standards (Table 3) simul-
taneously as leather samples, or when doubts arouse, proved to be
helpful to determine and compare the colour obtained in performed
tests.
The key-guide presented in Fig. 4 summarizes spot test results
possibilities and, in authors’ opinion, works as a useful tool to make
tannin type identification easier.
5. Conclusions
Chemical spot tests described in this work allowed tannins char-
acterisation in most of the tested samples, particularly in new
leathers, even when mixtures of tannins were used.
The adaptation of acidified butanol and nitrous acid tests, and
also the combined evaluation with rhodanine test, made possible
a more specific detection of the chemical nature of tannins used to
manufacture leather, overcoming limitations of the generally used
ferric and vanillin tests. With this set of tests it is now possible to
detect the three classes of vegetable tannins – gallotannins, ellag-
itannins and condensed tannins – in leather fibres and, indirectly,
to know roughly the vegetable sources used both in the past and
nowadays to produce leather.
It is not unusual that even leathers supplied for conservation
purposes, which claim to be tanned with hydrolysable tannins,
were found to contain condensed tannins. To avoid costly re-
restorations, leather conservators often need to quickly confirm
the technical characteristics provided by manufacturers of new
commercial leathers they are buying.
Since spot tests are a rapid, inexpensive and easy to perform
tool, in our opinion, they are particularly useful to be used by con-
servators in conservation ateliers.
Acknowledgements
Lina Falcão is grateful to Dr. Andreas Schulze, Landensamt für
Denkmalpfleg Sachsen of Dresden, for providing NL2 and HL2 sam-
ples, to Dr. Bernhard Trommer, Forschungsinstitut für Leder und
Kunstsoffbahnnen of Freiberg, for supplying HL1 sample and to
Katia Bittencourt, organic materials conservator, for supplying HL1
sample. Authors are grateful to the director and curator of the
National Palace of Ajuda (Lisbon, Portugal), Dr. Isabel Silveira God-
inho and Dr. Maria do Carmo Rebello de Andrade, respectively, for
permission to collect leather fibres identified as HL3.
References
[1] R. Thomson, Leather, in: E. May, M. Jones (Eds.), Conservation Science: Heritage
Materials, The Royal Society of Chemistry, Cambridge, 2006, pp. 92–119.
[2] A.D. Covington, Tanning Chemistry: the Science of Leather, The Royal Society
of Chemistry, Cambridge, 2009.
[3] R. Reed, Ancient Skins, Parchments and Leathers, Seminar Press, London, 1972.
[4] H. Kühn, Conservation and Restoration of Works of Art and Antiquities, Vol. I,
Butterworth, London, 1986.
[5] E. Haslam, Vegetable tannins, in: Encyclopedia of Life Sciences, John Wiley &
Sons, 2001, wiley.com/10.1038/npg.els.0001913.
156 L. Falcão, M.E.M. Araújo / Journal of Cultural Heritage 12 (2011) 149–156
[6] L.A. Clarkson, Developments in tanning methods during the Post-Medieval
Period (1500–1850), in: R. Thomson, J.A. Beswick (Eds.), Leather Manufacture
Through the Ages, Proceedings of the 27th East Midlands Industrial Archaeol-
ogy Conference, 15th October 1983, Northampton, 1983, pp. 11–21.
[7] B. Sol, L’évolution des procédés de tannage végétal, Environnement et
conservation de l’écrit, de l’image et du son, Actes des 2e Journées Inter-
nationales d’Études de l’ARSAG, 16 au 20 mai 1994, Association pour la
Recherche Scientifique sur les Arts Graphiques-ARSAG, Paris, 1994, pp. 89–
95.
[8] R. Córdoba de la Llave, Mil años de trabajo del cuero, Actas del II Simposium
de Historia de las Técnicas, 6-8 de Mayo de 1999, Ediciones Litopress, Córdoba,
2003.
[9] J. Waterer, John Waterer’s guide to leather conservation and restoration
(revised and abridged), The Museum of Leathercraft, Northampton, 1986.
[10] G.W.R. Ward (Ed.), Leather, The Grove Encyclopedia of Materials and Tech-
niques in Art, Oxford University Press, New York, 2008, pp. 338–342.
[11] R. Larsen, The chemical degradation of leather, Chimia 62 (11) (2008) 899–902.
[12] T. Sturge, The Conservation of Leather Artefacts, The Leather Conservation Cen-
tre, Northampton, 2000.
[13] M. Kite, R. Thomson, A. Angus, Materials and techniques: past and present,
in: M. Kite, R. Thomson (Eds.), Conservation of Leather and Related Materials,
Butterworth Heinemann-Elsevier, Oxford, 2006, pp. 121–129.
[14] R. Thomson, Towards a longer lasting leather: a summary of the CRAFT Leather
Project, Leather Conservation News 17 (1) (2001) 1–9.
[15] K. Khanbabaee, T. van Ree, Tannins: classification and definition, Nat. Prod. Rep.
18 (2001) 641–649.
[16] W. Vermerris, R. Nicholson, Phenolic Compound Biochemistry, Dordrecht,
Springer, 2006.
[17] F.L. Hilbert, Tanning materials (vegetable), in: R.E. Kirk, D. Othmer (Eds.), Ency-
clopedia of Chemical Technology, Vol. 13, The Interscience Encyclopedia, New
York, 1954, pp. 578–586.
[18] J.-P.H. Azéma, Moulins du cuir et de la peau : moulins à tan et à chamoiser en
France XIIe –XXe siècle, Éditions Créer, Nonette, 2004.
[19] A.D. Covington, The chemistry of tanning materials, in: M. Kite, R. Thomson
(Eds.), Conservation of Leather and Related Materials, Butterworth Heinemann-
Elsevier, Oxford, 2006, pp. 22–35.
[20] R. Larsen, Summary, discussion and conclusion, in: R. Larsen (Ed.), STEP Leather
Project: Evaluation of the Correlation between Natural and Artificial Ageing of
Vegetable Tanned Leather and Determination of Parameters for Standardiza-
tion of an Artificial Ageing Method – Research Report no. 1, The Royal Danish
Academy of Fine Arts, Copenhagen, 1994, pp. 165–180.
[21] H.R. Procter, The Making of Leather, University Press, Cambridge, 1914.
[22] J.C. Bickley, Vegetable tannins, in: C. Calnan, B. Haines (Eds.), Leather, its Compo-
sition and Changes with Time, The Leather Conservation Centre, Northampton,
1991, pp. 16–23.
[23] E. Haslam, Vegetable tannins – Lessons of a phytochemical life, Phytochemistry
68 (2007) 2713–2721.
[24] M. König, E. Scholz, R. Hartmann, W. Lehmann, H. Rimpler, Ellagitannins
and complex tannins from Quercus petraea bark, J. Nat. Prod. 57 (10) (1994)
1411–1415.
[25] M.-L. Florian, Vegetable tannins, Leather Conservation News 1 (4) (1984) 37–38.
[26] A. Pizzi, C. Simon, B. George, D. Perrin, M.C. Triboulot, Tannin antioxidant char-
acteristics in leather versus leather light stability: models, J. Appl. Polym. Sci.
91 (2004) 1030–1040.
[27] T.C. Smout, Oak as a commercial crop in the eighteenth and the nineteenth
centuries, Botanical J. Scotland 57 (1) (2005) 107–114.
[28] E. Cadahía, S. Varea, L. Muñoz, B.F. de Simón, M.C. García-Vallejo, Evolution
of ellagitannins in Spanish, French, and American oak woods during natural
seasoning and toasting, J, Agric. Food Chem. 49 (2001) 3677–3684.
[29] R. Larsen, J. Wouters, C. Chahine, P. Brimblecombe, C. Calnan, Recommenda-
tions on the production, artificial ageing, assessment, storage and conservation
of vegetable tanned leather, in: R. Larsen (Ed.), Environment Leather
Project: Deterioration and Conservation of Vegetable Tanned Leather-Research
report no. 6, The Royal Danish Academy of Fine Arts, Copenhagen, 1996,
pp. 189–202.
[30] C. Barbe, T. Aubry, T.-P. Nguyen, F. Juchauld, La sélection d’un cuir de tannage
végétal destiné à des traitements de restauration de livres anciens: une étude
en cours à la Bibliothèque nationale de France (BnF), Int. Preservation News 39
(2006) 4–8.
[31] H.-R. Tang, R.A. Hancock, A.D. Covington, Study on the composition and struc-
ture of commercial chestnut tanning agent, in: R.W. Hemingway, P.E. Laks
(Eds.), Plant Polyphenols: Synthesis, Properties, Significance, Plenum Press,
New York, 1992, pp. 221–243.
[32] C. van Driel-Murray, Practical evaluation of a field test for the identification of
ancient vegetable tanned leathers, J. Archaeol. Sci. 29 (2002) 17–21.
[33] D.V. Poulsen, Presentation and evaluation of spot tests for identification of
the tannin type in vegetable tanned leather, in: R. Vontobel (Ed.), Preprints of
ICOM Committee for Conservation, 13thTriennal Meeting, Vol. II, Rio de Janeiro,
22–27 September 2002, James and James Ldt, London, 2002, pp. 792–797.
[34] J. Wouters, High-performance liquid chromatography of vegetable tannins
extracted from new and old leathers, 10th Triennal Meeting, Washington,
22–27 August 1993, in: Preprints of ICOM Committee for Conservation, Vol.
II, James, James for ICOM-CC, London, 1993, pp. 669–673.
[35] R. Thomson, Testing leathers and related materials, in: M. Kite, R. Thomson
(Eds.), Conservation of Leather and Related Materials, Elsevier Butterworth-
Heinemann, Oxford, 2006, pp. 58–65.
[36] T. Klokkernes, Skin Processing Technology in Eurasian Reindeer Cultures, PhD
Thesis, The Royal Danish Academy of Fine Arts, School of Conservation, 2007.
[37] N. Odegaard, S. Carroll, W. Zimmt, Materials Characterisation Tests for Objects
of Art and Archaeology, Archetype Publications, London, 2000.
[38] B.H. Stuart, Analytical Techniques in Materials Conservation, John Wiley & Sons,
Chichester, 2007.
[39] A. Hagerman, Tannin chemistry, http://www.users.muohio.edu/hagermae/
tannin.pdf.
[40] A. Scalbert, Quantitative methods for the estimation of tannins in plant tissues,
in: R.W. Hemingway, E. Laks (Eds.), Plant Polyphenols: Synthesis, Properties,
Significance, Plenum Press, New York, 1992, pp. 259–278.
[41] P.C.M. Jansen, D. Cardon (Eds.), Plant Resources of Tropical Africa 3: Dyes and
Tannins, PROTA Foundation-Blackhuys Publishers-CTA, Wageningen, 2005.
[42] P. Schofield, D.M. Mbugua, A.N. Pell, Analysis of condensed tannins: a review,
Anim. Feed Sci. Technol. 91 (2001) 21–40.
[43] T.C. Wilson, A.E. Hagerman, Quantitative determination of ellagic acid, J. Agric.
Food Chem. 38 (8) (1990) 1678–1683.
[44] K.H. Inoue, A.E. Hagerman, Determination of gallotannins with rhodanine, Anal.
Biochem. 169 (1988) 363–369.
[45] Z.A. Kuliev, A.D. Vdovin, N.D. Abdullaev, A.B. Makhmatkulov, V.M. Malikov,
Study of the catechins and proanthocyanidins of Quercus robur, Chem. Nat.
Comp. 33 (6) (1997) 642–652.