Greener approach to leather tanning process: D-Lysine aldehyde as novel tanning

agent for chrome-free tanning

Ganesan Krishnamoorthy, Sayeed Sadulla, Praveen Kumar Sehgal
y
, Asit Baran Mandal
*
Council of Scientic & Industrial Research (CSIR), Central Leather Research Institute (CLRI), Adyar, Chennai 600 020, Tamil Nadu, India
a r t i c l e i n f o
Article history:
Received 12 December 2011
Received in revised form
29 October 2012
Accepted 2 November 2012
Available online 23 November 2012
Keywords:
Green chemistry
Greener process
Cleaner process
D-Lysine aldehyde
Chrome-free tanning
a b s t r a c t
A greener and eco-friendly method to leather tanning process has been developed in the present
investigation, which is based on D-Lysine aldehyde complex. This can be considered as a substitute for
chrome-free tanning process. The physico-chemical and morphological characteristics, resistance to
collagenolytic activity and organoleptic properties of the D-Lysine glutaraldehyde tanned leather were
evaluated in terms of environmental impact assessment. The shrinkage temperature, denaturation
temperature and mechanical strength of tanned leather were analysed by using shrinkage, differential
scanning calorimetric and thermomechanical analysis techniques. It has been shown that the above
properties of the tanned leather are better than that of glutaraldehyde alone or equal to chrome tanned
leather. The density of the bre structure throughout the cross section indicates the even structure and
bre bundles seem to be less dispersed compared to glutaraldehyde alone; were analysed by using
scanning electron microscopic and atomic force microscopic techniques. This tanning tanned leather
demonstrated a higher resistance to collagenolytic activity than conventional tanned leather. The tanned
leather exhibited good fullness, softness, smoothness, colour and general appearance compared to
glutaraldehyde alone. The signicant reduction in total solids content such as dissolved solids and
suspended solids present in the efuent compared to conventional tanning method is evident from
environmental impact assessment studies. This tanning process has the cost advantages due to the
possible decrease in chemicals consumptions and net saving from the reduced efuent treatment costs.
The improved tanning process reduces generation of toxic waste compared to chrome tanning process
and can be regarded as eco-friendly, which reducing the environmental impacts and contributes to the
development of greener or cleaner process.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
With the improvement of environment realisation and change
of consumption concept and manufacture patterns, the world
industry is keen to take on greener or cleaner approaches to leather
processing. Therefore, the researchers are compelled to explore
eco-friendly products and processes for the alternative chrome
tanning (Schramm, 1997). The leather tanning primarily employs
inorganic tannins viz., chromium, aluminium, ferrous and zirco-
nium; (Brown and Taylor, 2003) organic tannins such as, vegetable
tannins (Mavlyanov et al., 2001); aldehydes; synthetic and above
combinations are usually the conventional tanning methods
(Covington, 2008, 2009). The advantage of tanning, which improves
the robustness and viability of leather products, also prevents decay
and resistance towards chemical, thermal and microbiological
degradations. However, the disadvantages of the known chrome
tannins are limited available source, poor exhaustion and minimal
recoverable or reusable from leather wastes (Covington, 2009). The
vegetable tanning is time consuming, worker intensive, more
expensive and low quality product. The chrome tanning ensures
a maximum of 55e65% exhaustion of chrome resulting in wastage
as well as adding to pollution load (Covington, 2009). Further
disadvantages, chromium and other inorganic tannins include the
complicated process for waste treatment with net increase in
biochemical oxygen demand (BOD), chemical oxygen demand
(COD), total solid (TS), dissolved solids (DS), suspended solids (SS),
chromium and salts content (Tariq et al., 2009; Joseph and Nithya,
2009). The expensive disposal of leather scraps such as chrome
shavings, splitting, trimming and bufng dusts by dumping process
are also additional disadvantages (Hu et al., 2011). The pollution
problems of trivalent chromium (Cr(III)) in waste are due to
* Corresponding author. Tel.: 91 44 24910846/0897; fax: 91 44 24912150.
E-mail addresses: abmandal@hotmail.com, abmandal@clri.res.in, directorclri@
gmail.com (A.B. Mandal).
y
Deceased in October, 2011.
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Journal of Cleaner Production
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0959-6526/$ e see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jclepro.2012.11.004
Journal of Cleaner Production 42 (2013) 277e286
insufciency of the treatment systems and formation of hexavalent
chromium (Cr(VI)), carcinogenic and mutagenic, which led to the
search for an eco-friendly greener option to tanning process
(Kolomaznik et al., 2008).
Therefore, the combination tanning methods were considered as
suitable to overcome the above problems those arise from single
tanning process. The chrome-less and chrome-free combination
tanning processes are followed presently by the tanning industries.
For example, less chrome combination tanning includes chromium
with silica, chromium with iron, etc. The chrome-free combination
tanning based on organic such as, vegetable with synthetic tannin
and biocatalysis and inorganic such as, metal ions other than chro-
mium have been reported (Liu et al., 2006; Covington, 2008, 2009;
Kanth et al., 2009). Using these combinations of tannins in the
leather processing cannot achieve the characteristic properties, such
as, high performance quality and variety of leathers and their prod-
ucts. Therefore, for economic and ecological reasons the chrome
tanned leather must be considered extremely high so that no other
tanning processes have yet been found to provide the same univer-
sality, quality and variety of styles. However, all the above combi-
nation tanning do not have protable importance due to processing
complication, harmful effect and cost effect, etc. In the present
conditions, reduction of environmental impact of chromium and
other inorganics give extensive importance in the tanning process.
The cleaner process to chrome-free tanning, various small
molecules viz. urea, melamine, phenol, formaldehyde, and its
condensed products on the stability of the collagen and leather
tanning were analysed for reducing pollution loads (Simon and
Pizzi, 2003; Hui et al., 2011). The effects of urea and surfactant
(Rose and Mandal, 1996), and salt additive environments for micelle
of synthetic tanning materials (Mandal et al., 1981, 1987) and in the
stability, conformationand geometry of the collagenwere discussed
in detail in the past using hydrodynamic and thermodynamic
studies. These molecules attribute high tensile strength, elongation
modulus, low water absorption, mould shrinkage, high surface
hardness, elongation at break and volume resistance (Tillet et al.,
2011). Moreover, when these molecules and their derivatives are
used as material process, which may pose occupational hazardous
and harmful to human health (Cao et al., 2009).
Green chemistry or cleaner process in the sustainable devel-
opment is the design or re-design of chemical products and
processes that diminish or eliminate the use and generation of
hazardous substances, materials and also to be safer of chemical
transformations for human health and environmental friendly
(Manley et al., 2008; Bonilla et al., 2010; Klemes et al., 2012). The
introduction of green chemistry can not only help industry to
reduce its environmental, economic and social impact but could
also impart unexpected functional features such as thermo-
mechanical properties, proteolytic and microbial resistance
(Willems and Moens, 2010). The execution of green chemistry
approaches into chrome-free tanning could reduce the environ-
mental impact and thereby expected to yield a novel substitute
process for conventional tanning.
In order to improve the physico-chemical stability, organoleptic
properties and resistance to proteolytic activity, herein we have
studied tanning process using the unnatural D-amino acids viz. D-
form or mirror image of L-Lysine (Scheme 1) in the presence of
aldehyde. D-Lysine aldehyde tannedleather obtainedbythe reaction
of offered D-Lysine with aldehyde is alternative to aldehyde alone.
The incorporation of unnatural amino acids into proteins is an
important tool for understanding proteinfunctionsuchas structural
support, engineering proteins and introducing useful building
blocks and molecular scaffold for protein-based heterochiral
biomaterials (Annavarapu and Nanda, 2009). Reductions of the
environmental impact of chrome and alternative emerging options
to cleaner chrome tanning by exhaust aids using small molecules
(Sundar et al., 2002), oxalolidine (Li et al., 2009; Sundarapandiyan
et al., 2011) and tanning oats (Morera et al., 2011) for higher
chrome exhaustions have been studied. Recently, we have also
studied the green chemistry approaches to chrome-free tanning
process by unnatural D-amino acids viz. D-Alanine, D-Glutamic acid
and D-Lysine (Krishnamoorthy et al., 2012a,b,c). The report seeks to
overcome the above disadvantages of the existing tanning by using
unnatural D-amino acids with aldehyde as a tanning agent to
minimise the environmental impact of chromeintannerydischarge.
In this work, to reduce the environmental impact, cleaner
tanning process based on optically active unnatural D-Lysine (dia-
mino compound) with Glutaraldehyde (GTA) has been made as an
alternative chrome-free tanning with the improved physico-
chemical and organoleptic properties. The tanning conditions of
D-Lysine GTA, physico-chemical characteristics of leathers and the
environmental impact assessment and economic analysis of this
tanning method have been compared with conventional tanning
process with a view to stimulate further studies.
2. Experimental methods
2.1. Materials
The raw material of pickled pelt processed from wet salted goat
skin was used for tanning processing. All the chemicals were
commercial grade, whichare generally usedinthe leather industries.
2.2. Preparation of D-Lysine
The production of D-Lysine by racemisation was done using the
following modied method (Takahashi et al., 1997). The race-
misation of L-Lysine was performed using a 200 mL solution of 30%
L-Lysine hydrochloride in a 300 mL autoclave was stirred at 135

C
for 105 h. To the resulting solution, 800 mL ethanol was added to
crystallise-out D-Lysine. The yield of D-Lysine was 90%.
2.3. Leather tanning process
The tanning process for both control and experiments were
carried out using pickled goat skins and the cumulative input of
tanning process was presented in Table 1. Two set of three pickled
NH
2
O
OH
H
2
N
NH
2
O
OH
H
2
N
L-Lysine
D-Lysine
Scheme 1. Proposed structure of stereochemistry of L-Lysine and D-Lysine.
G. Krishnamoorthy et al. / Journal of Cleaner Production 42 (2013) 277e286 278
goat skins were treated with 0.5% sodium bicarbonate for depick-
ling and than tanned by 1% D-Lysine and 2% Glutaraldehyde (GTA)
based on pickled weight, for 3 h until penetrated at pH 3.5e4.5 and
then the xing was carried out by the addition of 1% formic acid and
4% Basytan FB 6 in four instalments at an interval of 15 min with
little modication compared to our recent report (Krishnamoorthy
et al., 2012a). The control process was carried out without treating
the pelt with D-lysine and with 11% Basyntan FB 6. Finally, the
tanning drum was run for 1 h. The tanned leathers were removed
from the tannage bath without washing, piled and dried. The
physico-chemical properties of the leathers obtained were
assessed. The control and experimental tanning process liquor from
the each set of trial has been collected for the environmental
impact assessment analysis.
2.4. Physical testing
2.4.1. Shrinkage temperature
The shrinkage temperature of D-Lysine GTA tanned leather was
measured using a micro-shrinkage tester using the standard IULTCS
method. A small strip was cut from the tanned leather and placed
on a water-grooved microscope slide, which in turn was placed on
a heating stage along with a microscope mounted above the
heating stage. The rate of heating was maintained at 2

C/min. The
temperature at which the leather bre shrinks to one-third of its
original length was taken as the shrinkage temperature.
2.4.2. Mechanical strength
The tensile strength and % elongation have been examined using
the standard IULTCS methods. The mechanical properties of D-
Lysine GTA tanned leather were analysed using Thermomechanical
analyser (TMA, Instron series II Automated Materials Testing
System). The mechanical properties were assessed in comparison
with conventional method, after conditioning the crust leather at
25

C with relative humidity over a period of 48 h.
2.4.3. Denaturation temperature
The differential scanning calorimetric (DSC, TA-DSC Q 200)
technique was used for analysis of denaturation temperature of D-
Lysine GTA tanned leather. The sample was weighed (5 mg) into
a hermetic aluminium pan, which was then sealed using a sample
pan crimper. The thermogram was obtained by heating to 25e
300

C at the rate of 10

C per min. The reference cell used in the
analysis was an empty hermetic aluminium pan. The standard
Indium was used for calibration of the temperature scale. The
denaturation temperature of the tanned leather was obtained from
the peak temperatures.
2.4.4. Morphological analyses
The scanning electron microscopic (SEM, FEI QUANTA 200) and
atomic force microscopic (AFM, Park Systems XE-70) techniques
were used for analyses of morphological characterisations of D-
Lysine GTA tanned leather. Sample from tanned leather was cut
from the ofcial sampling position. The specimen was cut with
uniform thickness without any pre-treatment. The micrographs for
the grain surface and cross section were obtained by operating the
SEM. The micrograph for the grain surface was also obtained by
using AFM.
2.5. Organoleptic properties
The matched pair was assessed for softness, fullness, grain
atness, grain smoothness, grain tightness (break) and general
appearance of D-Lysine GTA tanned leather by hand and visual
examinations. The tanned leather was rated (on a scale of 1e10
where 1 is poorest and 10 is the best) for each functional prop-
erty by experienced tanners.
2.6. Biodegradation
The percentage of degradationof the D-Lysine GTAtanned leather
against collagenase activity was calculated by using earlier modied
method described recently in details (Krishnamoorthy et al., 2011).
The tanned leather was treated with collagenase. The tanned
leather: collagenase ratio (25 mg tanned leather: 0.5 mg collage-
nase) was maintained at 50:1. The sample was incubated at
a temperature of 37

C. Sample was collected at various time inter-
vals ranging from 6 to 96 h and stored in freezer. The cleavage of
tanned leather was monitored by the release of soluble form of
hydroxyproline from insoluble collagen. Aliquot of 750 mL of super-
natant was withdrawn after centrifuging at 10,000 rpm for 10 min.
The collagenase hydrolysate was hydrolysed by 6 N HCl in sealed
hydrolysis tubes for 16 h. The nal hydrolysate was evaporated to
dryness in a porcelain dish over a water bath to remove excess acid.
The residues of free acid were made up to a known volume and the
percentage of hydroxyproline was determined by using this method
(Woessner, 1961). Hydroxyproline is an unique amino acid for
collagen and offers a useful marker for identifying collagen in
presence of non-collagenous proteins. The method of determining
hydroxyproline involves the oxidation of hydroxyproline to pyrrole-
2-carboxylic acid, which complexes with p-dimethylamino-
benzaldehyde exhibiting maximum absorbance at 557 nm.
% Soluble collagen % Hydroxyproline 7:4
The % degradation of collagen for D-Lysine GTA tanned leather is
calculated based on the soluble collagen content in the supernatant
solution of the collagenase treated D-Lysine GTA tanned leather.
2.7. Environmental impact assessment
The biochemical oxygen demand (BOD) and chemical oxygen
demand (COD) were analysed by using the standard procedure
(APHA, 1995) for the D-Lysine GTA tanned exhaust liquor from
experimental processing. The total solids (TS) content, dissolved
solid (DS) and suspended solids (SS) were calculated. The values
reported here are the averages of 3 experiments along with their
standard errors (SE). A complete inputeoutput for the raw mate-
rials, water, chemicals and cost effect of chrome, GTA and D-Lysine
GTA tanning processes for 1 kg of Indian goat skins were analysed.
2.8. Statistical analysis
Data are expressed as the mean standard error (SE). Statistical
analysis was performed using the Students t-test.
3. Results and discussion
The improved protein stability can be achieved by incorporating
variable unnatural amino acids of D-conguration stereochemistry
in molecular devise and create newtopologies inaccessible to homo
Table 1
Cumulative input of chemicals for D-Lysine GTA tanning process.
Processes % water Sodium
bicarbonate
% D-Lysine pH % GTA % water Formic
acid
Basytan
FB 6
GTA 2.8 11%
D-Lysine
GTA
50 0.5% 1 4.4 2 50 1% 4%
G. Krishnamoorthy et al. / Journal of Cleaner Production 42 (2013) 277e286 279
chiral molecules (Annavarapu and Nanda, 2009). The unnatural
stereochemistry often means that molecules containing D-residues
are much more resistant to enzyme-catalysed breakdown and
antimicrobial activity than natural peptides, a property of consid-
erable pharmaceutical and microbiological importance from
a functional point of view (Oh and Lee, 1999). Schematic repre-
sentation of collagen side chain crosslinking with D-Lysine GTA
leading to formation of amide bond is depicted in Schemes 2 and 3.
The effect of the D-Lysine GTA used as a novel tanning agent on
enhancing the mechanical strength, hydrothermal stability and
resistance to the collagenolytic biodegradation are presented here.
As expected, reduced environmental and economic impact such as
biochemical oxygen demand (BOD), total solids (TS), dissolved
solids (DS), suspended solids (SS) and cost effect for the tanned
leather can be gained when the tanning is conducted in the pres-
ence of D-Lysine GTA.
3.1. Mechanism of action of D-Lysine GTA
The D-Lysine GTA can combine with collagen and improve the
hydrothermal stability of the tanned leather. The amino groups of
collagen side chains after combining with the D-Lysine GTA become
unavailable for hydrogen ions. This indicates the mechanism by
which D-Lysine GTA combines with collagen (Schemes 2 and 3). Its
reaction with collagen is commonly assumed to involve the
formation of Schiff bases between the two carbonyl ends of GTA
and positively charged amino groups on the surface of the collagen
and offered D-Lysine. Research has indicated the best candidate to
be the -amino group of lysine, hydroxylysine and arginine side
chains of collagen in the protein molecules creating a structure
more stable than other proteins. In process, GTA easily polymerises
by condensation, giving rise to mixtures of elongated species that
can in turn also cross-link intra and intermolecular lysine of
collagen in a nonspecic manner. The additional and surprising
ring formation on the arginine side with GTA could arise from the
formation of a further Schiff base between one of the nearest
-aldehyde groups of the GTA monomer/oligomer and the second
amino group of arginine. Revisiting GTA cross-linking: the case of
the arginineelysine intermolecular doublet has been studied
(Salem et al., 2010). The previously exposed amino groups facilitate
in increasing the stability of tanned leather. This explains the
increase in uptake of D-Lysine when GTA is offered prior to GTA
tanning. Interestingly, the D-Lysine uptake was found to be more
when GTA was offered. Amino groups of both lysine and hydrox-
yproline are involved in the GTA tanning reaction. Proximity of the
amino groups to the GTA is much essential for GTA tanning and
improves the properties. It may be possible that D-Lysine might
have improved the hydrothermal stability of tanned leather.
3.2. Hydrothermal stability
The denaturation temperature of D-Lysine GTA tanned leather
was studied employing DSC. The thermal denaturation of leather
was obtained over a dened temperature range by considering the
position, width, height and symmetry of the thermogrampeak. The
hydrothermal stability of tanned leather subjected to D-Lysine GTA
treatment was normally related to the onset temperature of peak in
DSC (Fig. 1A and B). An increase in the denaturation temperature
93 3 to 108 7

C is usually an indication of an increase in the
stability of wet tanned leather. Fortunately, in case of hide treated
with D-Lysine GTA tanned leather become narrow and sharp,
moreover, the shrinkage temperature for GTA and D-Lysine GTA of
nal leather reach 90 4 and 120 7

C, respectively (see Table 2).

The D-Lysine GTA tanning effect is superior to GTA alone, which has
been demonstrated with improved hydrothermal stability of the
leather. The shrinkage temperature of the leather was estimated to
ascertain the GTA level in the leather and the extent of cross links
formed during tanning. The D-Lysine GTA tanned leather was found
to be soft and full possessing tight grain. This may be due to the
decrease in the availability of the active sites in the leather matrix.
The hydrothermal stability of brillar collagens depends on intra
Scheme 2. Schematic representation of proposed structure of side chain crosslinking
of collagen with D-Lysine GTA. The result of crosslinking bridge via amino group of two
collagen lysine side chain are directly joined via an amide bond and also play the major
role in stabilisation of collagen in presence of GTA.
Scheme 3. Schematic representation of proposed structure of side chain crosslinking
bridge of collagen with D-Lysine GTA. The result of crosslinking bridge via amino group
of collagen lysine and arginine side chain are directly joined via an amide bond and
also play the major role in stabilisation of collagen in presence of GTA. The arginine
side could arise from the formation of a further Schiff base between one of the nearest
-aldehyde groups and the second amino group of arginine.
G. Krishnamoorthy et al. / Journal of Cleaner Production 42 (2013) 277e286 280
and intermolecular cross-links formed through the D-Lysine GTA.
This indicates clearly that the treatment using 1% D-Lysine facili-
tates offer of 2% GTA that inuence the quality of the leather.
3.3. Mechanical properties
It is essential to analyse the mechanical strength properties
upon treatment with D-Lysine GTA. The D-Lysine GTA tanned
leather tensile strength and % elongation at break were determined
and compared with GTA tanned leather (Table 2). The above tanned
leather is very close to the BCS tanned leather. The tanned leather
exhibited high tensile strength and low % elongation, whereas GTA
tanned leather exhibited low tensile strength with high % elonga-
tion. The tanned leather shows cemented bres bundles, whereas
GTA alone tanned leather bre bundles are well separated, as
revealed by SEM studies (Fig. 2B). It is seen that experimental
leather exhibit higher mechanical strength values compared to
control leather.
3.4. AFM analysis of grain surface roughness
The plane and three-dimensional proles of the grain surface of
D-Lysine GTA tanned leather are shown in Fig. 2A. The lighter and
darker areas of the images correspond to higher and lower topog-
raphies, respectively. The tanned leather shows a rougher texture
with higher peaks than the GTA tanned leather. Tanning by D-Lysine
GTA is thought to be the reason for the increased stability of the
tanned leather, which is manifested by higher hydrothermal
stabilities. The presence of D-Lysine GTA clearly formed the bres in
the AFM images where the typical collagen banding pattern of
grooves and ridges are obtained (Fig. 2A).
3.5. SEM analysis
The D-Lysine GTA tanned leather sample showing grain surface
using SEM is shown in Fig. 2B. There is no physical deposition,
which is evident from a clear grain surface obtained in the tanned
leather. Higher magnication (1000) SEM (Fig. 2C) conrms the
above observation. The hair follicles look clean without any foreign
materials in all cases. However, the grain surface of the tanned
leather seems to be at without any wrinkles compared to the
leather tanned by GTA alone. The cross section of tanned leather at
a magnication of 500 is depicted in Fig. 2D. The density of the
bre structure throughout the cross section indicates the even
structure of the tanned leather. The tanned leather sample of the
bre bundles seem to be less dispersed compared to GTA alone. In
principle, increased fullness and softness of the D-Lysine GTA tan-
ned leather exhibit slightly dispersed bre structure.
3.6. Organoleptic properties
The organoleptic properties of D-Lysine GTA tanned leather are
given in Table 3. From the Table 3, it is observed that the tanned
leather exhibited the organoleptic properties viz. good fullness,
softness, smoothness, colour and general appearance compared to
GTA alone. Fullness and tightness were found to be better when D-
Lysine was offered before tanning. It was found that the
enhancement of fullness and tightness was directly proportional
to the D-Lysine offer. This is because of the improved structural
properties of D-Lysine GTA tanned leather. Generally, the appear-
ance and overall performance of the tanned leather is good
compared to GTA alone. From the results, it is evident that the
offer of 1% of D-Lysine would be optimum to accrue the benecial
effects in terms of leather properties. Given the signicant
improvement in fullness and tightness, washing and persist due to
the treatment using D-Lysine GTA, it was constructed that there is
a possibility of reduction in the syntans during retanning.
However, the fullness and tightness of the leather were enhanced
due to D-Lysine GTA treatment, softness was found unaltered.
Therefore, there was no need for additional amount of fatliquor.
Thus, the experiment was conducted converting the convention-
ally GTA tanned leather treated with 1% D-Lysine and 2% GTA into
crust leathers using lower amount of syntans. The cumulative
input of chemicals for D-Lysine GTA tanning process is given in
Table 1. The crust leather was tested for physical properties
(Table 2). It can be observed from the results that the physical
characteristics such as tensile strength and % elongation were
found to be better while D-Lysine GTA was offered.
3.7. Biodegradation
The D-Lysine GTA tanned leather degradations by collagenase
at different concentrations have been determined and shown in
Fig. 3A and B. The percentage reduction of degradation of leather
Fig. 1. DSC analysis of D-Lysine GTA tanned leather.
Table 2
Denaturation temperature (T
d
), shrinkage temperature (T
S
), tensile strength (TS) and
percentage of elongation (%E) of D-Lysine GTA tanned leather.
Process T
s
(

C) T
d
(

C) TS (MPa) % E
GTA 90 4 93 3 15 2 72 6
D-Lysine GTA 120 8 108 7 18 3 56 6
G. Krishnamoorthy et al. / Journal of Cleaner Production 42 (2013) 277e286 281
is observed signicantly for tanned leather compared to GTA
alone. The 9% degradation was observed in D-Lysine GTA tanned
leather against 99% degradation in case of untanned (depickled
skin) at 96 h period of incubation. The stability of D-Lysine GTA
tanned leather is obtained by protecting the active sites in
collagen through interaction with D-Lysine GTA not recognised by
collagenase. The degradation of D-amino acids protein and protein
based materials are not well equipped by the proteolytic
machinery, however, it is this result above all else that has
spurned research into stereochemical inversion and directional
manipulation of peptide and polypeptide chains. The hope has
been that organised inversion of the stereochemistry at the
peptide backbone a-carbon atoms, if accompanied by chain
U-turn, should yield proteolytically stable retro-inverso peptide
isomers (Fischer, 2003).
3.8. Environmental impact assessments
The environmental impact analysis of tanning process is
assessments of the possible positive or negative impact have on the
environmental, social and economic aspects. Biochemical oxygen
demand, chemical oxygen demand (COD) and total solids are main
parameters in assessing the quality of spent tan liquor. The spent
tan liquor analyses for biochemical oxygen demand (BOD), chem-
ical oxygen demand (COD), total solids (TS), dissolved solids (DS)
and suspended solids (SS) were assessed. It can be seen from
Table 4 that the COD value in combination tanning is equal to GTA
alone. However, the TS load value is lower than that of the GTA
tanning process. The pickling process increases the TS of the
efuent, which is a well known phenomenon. The pollution of
neutral salt is signicantly decreased in D-Lysine GTA tanning. The
BOD/COD value is greater for D-Lysine GTA tanning in comparison
with GTA tanning. The TS values for D-Lysine GTA tanned leather are
lower than that for the control leather processing, in spite of
reduced chemical consumption. The reductions in the BOD and TS,
DS and SS are primarily due to the modied tanning process, which
reduces the environmental impact. It is evident that the tanning
method reduces the BOD, TS, DS and SS loads by 55, 53, 60 and 75%,
respectively. This process eliminates the chrome, which leads to
signicant reduction in TS. The reduction in BOD, TS, DS and SS
helps in achieving reduction of environmental impact. Execution of
this tanning method could bring signicant change in the tanning
industry producing the experimental leather process to have the
advantage of zero release solid wastes.
Fig. 2. Morphological characterisation of D-Lysine GTA tanned leather: A) AFM image grain surface, B) SEM image grain surface, C) SEM image grain surface, D) SEM image cross
section. The collagen brils cross-linked with D-Lysine revealed a well-ordered structure with the brils properly oriented and well aligned.
G. Krishnamoorthy et al. / Journal of Cleaner Production 42 (2013) 277e286 282
3.9. Economic analysis
The execution of new cleaner process requires new commercial
possibility and cost efcacy. This process to be considered
sustainable developments must offering reduce environmental
impact and be economically viable. The total chemical costs for
processing 1 kg of goatskins through eld trials conventional
chrome, GTA and experimental process schemes are given in
Table 5. It could be seen that the experimental process exhibits
lowest chemical cost as compared to the control process. The GTA
tanned leather was converted into crust leather using syntan and
fatliquors offer similar to conventional process. The D-Lysine
assisted GTA tanned leather was converted into crust leathers using
syntans of reduced offer levels considering the enhancement in the
fullness and tightness due to offer of D-Lysine. However, the leather
did not become hard and the softness was found unaltered.
Therefore, the fatliquor offer level was similar to the control and the
syntan offer was reduced. For D-Lysine GTA treated leather, only 4%
of syntan was offered against 11% of syntan in control leather. The
offers of D-Lysine with GTA decrease the cost signicantly, for
processing 1 kg of raw goatskins. It could be seen that the D-Lysine
with GTA exhibits lower chemical cost as compared to the
conventional chrome and GTA tanning process. The total chemical
cost of the conventional chrome and GTA tanning processes are
about US$ 0.4074 and US$ 0.3137, respectively, whereas experi-
mental D-Lysine aldehyde tanning process is about US$ 0.2207.
Hence, the possible decrease in chemical cost is about US$ 0.1867,
for processing 1 kg of rawgoatskins. This improved tanning process
reduces the discharge of solid and liquid wastes, BOD, chromium,
chloride, TS, DS and SS loads which provide further protable in
efuent treatment costs compared to chrome tanning process. The
results indicate that the D-lysine GTA tanning is more economical
than the GTA alone. It must be pointed out that the adoption of this
tanning does not demand any kind of investment or additional
expenditure. This tanning method leads to a viable greener option
to the conventional intricate processing. Such greener option can
be followed in leather sector both developed and developing
countries as it has potential to reduce environment impact with
near zero pollution from tanning process without affecting the
organoleptic properties of leather.
Using this D-Lysine with GTA for tanning is an effective method of
eco-friendly processing. D-Lysine is a smaller molecule when offered
before GTA tanning was found to enhance the stability equal to
chromiumtannedleather. This canexplainthat D-Lysine canenhance
the mechanical strength, hydrothermal stability and resistance to
collagenolytic activity with improved property that is inaccessible to
proteolytic machinery. This D-Lysine GTAtanning studysuggests that
free amino groups exposed by collagen brils are initially reacted
with GTA and then bridged by the D-Lysine (diamino compound)
producing more extensive inter and intra molecular crosslinking
than GTA alone tanned leather. The schematic representation of
amide bondformed fromreactions between hide collagenlysine and
hydroxylysine side chain with L- and D-Lysine GTA is shown in the
Fig. 2. (continued).
Table 3
Rating of organoleptic properties of D-Lysine GTA tanned leather.
Processes Fullness Grain
tightness
Grain
smoothness
Softness Colour of
tanned leather
General
appearance
GTA 8.5 7 7.5 9 8 8
D-Lysine
GTA
9 8.5 9 9 8 8.5
G. Krishnamoorthy et al. / Journal of Cleaner Production 42 (2013) 277e286 283
Scheme 2. The collagen matrix largely relies on D-Lysine possessing
non-natural structures or a non-natural absolute conguration was
found not to inuence the leather quality negatively. D-Lysine is not
harmful to human health, evidenced from used in nutrient (Kano
et al., 2003) and used as supplementary material for variety of cell
cultures (Deng et al., 2011). The properties such as fullness and
tightness were found to be improved signicantly by D-Lysine indi-
cating the possibilityof reducingthe offer of syntanlater. As far as the
offer level is concerned, the optimum offer was found to be around
1e2%. D-Lysine GTA was found to be beneted from signicant
reductionintotal solidcontent andimprovedbetter biodegradability
of organic compound present in the efuent compared to conven-
tional tanning method. This improved greener or cleaner approach
for achieving zero discharge in the tanning industry has become
a reality.
4. Conclusions
The global warming or climate change issue is eager to adopt
greener or cleaner approach to leather, textile and paper process-
ing. The cleaner approach to chrome-free tanning based on D-Lysine
GTA has been developed. This optimised tanning process has been
adopted for employing a chrome-free tanning. The physico-
chemical properties of the leather are improved with respect to
texture, hydrothermal stability, mechanical strength, resistance to
collagenolytic activity, and also organoleptic properties. The
important advantages of the D-Lysine GTA approach is the unex-
pected environmental benets achieved. The D-Lysine GTA tanning
efuent shows reduction in BOD, TS, DS and SS as compared to
control process, apart from achieving better organoleptic proper-
ties. In general, this approach provides an abundant scope for
decreasing the pollution load either with regard to liquid waste or
solid waste. The economic protable analysis study reveals that the
D-Lysine GTA tanning process has the cost advantage due to the
possible decrease of the total chemical consumption and net saving
from the reduced efuent treatment cost compared to control and
chrome tanned leather processing. Further advantages of D-Lysine
aldehyde tanning include reducing the quantity of leather scraps
such as shavings, splitting, trimming and bufng dusts by dumping
process. This tanning process has numerous benets in terms of
reduction of environmental impact, improved leather quality with
better global reection. Additional advantages will also ensure for
the inhabitants living in the surrounding area of the beneciary
tannery units and the working employees as well as reduction in
Fig. 3. Biodegradation degree: A) GTA and D-Lysine GTA crosslinked tanned leather and B) various concentrations of D-Lysine with constant GTA. The values reported are average of
3 experiments along with their standard errors (SE). Statistically signicant (*p < 0.05, **p < 0.005, ***p < 0.0005).
Table 4
Biochemical oxygen demand (BOD), chemical oxygen demand (COD), total solid contents (TS), dissolved solid (DS) and suspended solid (SS) analyses of D-Lysine GTA tanned
leather.
Process pH COD (ppm) BOD (ppm) TS (ppm) DS (ppm) SS (ppm)
GTA 3.0e4.0 1200e3000 400e1000 20,000e40,000 25,000e48,500 800e1800
D-Lysine GTA 3.0e4.0 1200e3200 200e600 11,000e26,000 16,000e32,000 300e1200
Table 5
Economic analysis of chrome, GTA and D-Lysine GTA tanning processes: 1 kg of
Indian goat skins.
Process Chemicals Cost per kg
(US$)
Input
(kg)
Cost
(US $)
Chrome Sodium chloride 0.054 0.1 0.0054
Sulfuric acid 0.2727 0.01 0.0273
BCS 1.20 0.08 0.0960
Basytan FB 6 2.36 0.11 0.2596
Sodium formate 0.55 0.01 0.0055
Sodium bicarbonate 1.36 0.01 0.0136
Total 0.4074
GTA Sodium bicarbonate 1.36 0.005 0.0068
GTA 3.18 0.01 0.0318
Formic acid 1.55 0.01 0.0155
Basytan FB 6 2.36 0.11 0.2596
Total 0.3137
D-Lysine GTA D-Lysine (for cost of
synthesized D-Lysine,
costs of L-Lysine and
ethanol taken into account)
4.00 0.01 0.0400
Sodium bicarbonate 1.36 0.005 0.0068
GTA 3.20 0.02 0.0640
Formic acid 1.55 0.01 0.0155
Basytan FB 6 2.36 0.04 0.0944
Total 0.2207
Possible cost difference between D-Lysine GTA tanned leather
compared
to chrome tanning
0.1867
G. Krishnamoorthy et al. / Journal of Cleaner Production 42 (2013) 277e286 284
land and water pollution, better work environment and improved
work-related safety measures are the major benets. The adoption
of D-Lysine GTA tanning method would be regarded as eco-friendly,
comparable to chrome tanning process, which can contribute to the
reduction of environmental impact.
Acknowledgments
One of the authors (GK) acknowledges the Council of Scientic &
Industrial Research (CSIR), Government of India, New Delhi, for his
CSIR-SRF fellowship.
Glossary
AFM Atomic force microscopy
BCS Basic chromium sulfate
BOD Biochemical oxygen demand is the measure of the
degradable organic material present in a sample, and can
be dened as the amount of oxygen required by the
microorganisms in stabilizing the biologically degradable
organic matter under aerobatic condition.
COD Chemical oxygen demand is the measure of oxygen
consumed during the oxidation of the organic matter in
water by a strong oxidizing agent.
DS Dissolved solids
DSC Differential scanning calorimetric
D-Lysine D-Lysine is mirror image of L-lysine, which creates new
topologies inaccessible to homo chiral molecules and
resistance to proteolytic activity on protein.
EIA Environmental impact assessment, assessments of
possible positive or negative impact have on the
environmental, social and economic aspects.
GTA Glutaraldehyde
IULTCS International Union of Leather Technologists and
Chemists Societies
SEM Scanning electron microscopy
SS Suspended solids
Basyntan FB6 Synthetic tanning agent used to ll the voids of
leather matrix
T
d
Denaturation temperature
TDS Total dissolved solids, comprise inorganic salts and small
amounts of organic matter that are dissolved in water.
TMA Thermomechanical analysis
T
s
Shrinkage temperature
TS Total solids, which includes dissolved and suspended
solids.
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