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Physico-chemical characteristics of leaf litter biomass to delineate the

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DOI: 10.1016/j.jtice.2016.02.011

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 Journal of the Taiwan Institute of Chemical Engineers 62 (2016) 239–246

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Journal of the Taiwan Institute of Chemical Engineers

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

Physico-chemical characteristics of leaf litter biomass to delineate the

chemistries involved in biofuel production
Nadeem Akhtar a, Dinesh Goyal a,∗, Arun Goyal b
a
Department of Biotechnology, Thapar University, Patiala, 147 004 Punjab, India
b
Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, 781 039 Assam, India

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

Article history: Increase in energy demand across the world has put immense pressure on utilization of widely avail-
Received 10 September 2015 able lignocellulosic agricultural waste biomass and forest residues. Physico-chemical characteristics of leaf
Revised 2 January 2016
litter from largely grown tree species such as Mangifera indica, Populus deltoides and Polyalthia longifo-
Accepted 2 February 2016
lia were evaluated for possible use in biorefinery. Physical and chemical properties of these leaf litter
Available online 16 March 2016
biomasses were examined using bomb calorimetry, SEM, XRD, TGA, CHNSO analysis, FTIR and solid state
13
Keywords: C CP/MAS NMR spectroscopy. Low ash content (3.70 wt%), high volatile matter (76.05 wt%) and cellu-
Biofuel lose (37.75 wt%) was observed from leaf litter biomass of P. deltoides. SEM of leaf litter biomass revealed
Mangifera indica compacted surface morphology of M. indica, however a fibrillar structure was observed in P. deltoides and
Populus deltoides P. longifolia. Maximum crystallinity index (CrI) was observed in leaf litter biomass of M. indica (23.13%)
Polyalthia longifolia followed by P. longifolia (20.94%) and P. deltoides (20.93%). The calorific values of all biomasses were in
the range of 18.37 to 19.32 MJ/kg. Delineation of these physico-chemical characteristics together per se
shows that leaf litter biomass can also act as a potential feedstock for biofuel production.
© 2016 Taiwan Institute of Chemical Engineers. Published by Elsevier B.V. All rights reserved.

1. Introduction
Considering world energy demand and its security, attention is
being focused on inexpensive renewable lignocellulosic substrates
for biofuel production. These energy resources are an alternatives
to fossils fuel and their derivatives, and have become a high prior-
ity for chemical industries [1]. To solve the global warming prob-
lem and ensure sustainable development of the economy, it is nec-
essary to increase the use of renewable biomass resources [2], as
they lack the environmental risk and capital investment associated
with fossil fuels. The available biomass of the world is 220 billion
oven dry tones (ODT) per year or 4500 EJ (1018 J) [3]. India meets
70% of its energy requirement using fuel woods and in the process
about 50 million tones of wood are removed from forests every
year [4]. Substantial portion of cellulosic and hemicellulosic com-
ponent of leafy biomass can be readily hydrolyzed into fermentable
sugars by action of different hydrolytic enzymes, which is further
biotransformed into ethanol [5]. Therefore, leaf litter biomass from
Abbreviations: CrI, crystallinity index; CP/MAS NMR, cross polarizing magic an-
gle spinning nuclear magnetic resonance; FTIR, fourier transform infrared; SEM,
scanning electron microscopy; TGA, thermal gravimetric analysis; XRD, X-ray
diffraction.
∗
Corresponding author. Tel.: +91 175 2393314; fax: +91 175 2393011.
E-mail address: dgoyal@thapar.edu (D. Goyal).
tree plantation sites can be collected and used as a promising feed-
stock for biofuel production to mitigate energy crisis.
Lignocellulosic biomass refers to plant derived organic mat-
ter, composed of cellulose, hemicellulose, lignin, pectins, extrac-
tives, glycosylated proteins and several other inorganic mate-
rials. Cellulose is a linear condensation polymer consisting of
D-anhydroglucopyranose joined by β -1, 4-glycosidic bonds with a
degree of polymerization from 100 to 20,0 0 0 [6]. Hemicellulose,
a complex carbohydrate, is comprised of pentoses (e.g. xylose and
arabinose), hexoses (e.g. mannose, glucose and galactose) and sugar
acids. The most abundant polymer after cellulose and hemicel-
lulose is lignin, which consists of three different phenylpropane
units (p-coumaryl, coniferyl and sinapyl alcohol) [7]. The cellu-
lose, hemicellulose and lignin content of such biomasses fall in the
range of 30–50%, 15–35% and 10–20%, respectively [8,9]. The ac-
cessibility of cellulose which is embedded in hemicelluloses-lignin
matrix and highly crystalline nature of the cellulose are two main
factors responsible for increasing the cost associated with process-
ing of lignocellulosic biomass [10]. Determining relative quantities
of cellulose, hemicellulose and lignin in lignocellulosic feedstock
is often required in studies involving fractionation [11], fermenta-
tion [12], or other modification [13]. Besides biofuel, lignocellulosic
biomass can be a source of other bio-based products such as lev-
ulinic acid, furfural and hydroxy methyl furfural [14].
Chemical and molecular characteristics of lignocellulosic feed-
stock vary with geo-climatic conditions and taxonomic rank too.

http://dx.doi.org/10.1016/j.jtice.2016.02.011
1876-1070/© 2016 Taiwan Institute of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
240 N. Akhtar et al. / Journal of the Taiwan Institute of Chemical Engineers 62 (2016) 239–246
Table 1
Botanical nomenclature of biomass samples source.
Common name Family Genus Scientific name
Mango Anacardiaceae Mangifera Mangifera indica
Poplar Salicaceae Populus Populus deltoides
Ashoka Annonaceae Polyalthia Polyalthia longifolia
The primary obstacle impeding the widespread production of
bioenergy from biomass lies in the availability of low-cost tech-
nology to overcome recalcitrant nature of lignin. To make cellulose
more accessible for hydrolytic enzymes, lignin component must be
separated [15], which primarily depends upon the degree of poly-
merization, crystallinity, structural composition and surface area
[16]. Knowing accurately the physico-chemical composition of lig-
nocellulosic biomass is gaining importance in assessing its poten-
tial for biomass to biofuel conversion [1,11,17].
The current investigation is based on evaluation of physico-
chemical characteristics of different leaf litter biomasses from
largely grown tree species such as Mangifera indica, Populus del-
toides and Polyalthia longifolia to evaluate their potential as feed-
stock for bio-energy production.
2. Material and methods
2.1. Collection of biomass samples
Leaf litter biomass samples from largely grown trees such as M.
indica, P. deltoides and P. longifolia were collected from soil surface
in early spring (March 2014) from Thapar University campus, Pa-
tiala, Punjab (India), located at 30°19 48"N, 76°24 0"E and 310 m
above the sea level. The detailed nomenclature of the source of
biomasses is represented in Table 1. The air dried biomass sam-
ples were ground using mechanical blender and sieved to get ho-
mogeneous powder (diameter of particle < 0.5 mm) (Supplemen-
tary material, Plate 1). Sieved biomass samples were stored in air
tight containers at room temperature for characterization studies.
The general outline of the biomass characterization is presented in
Fig. 1.
2.2. Physical characterization
2.2.1. Moisture content
Moisture content plays a key role in the selection of a suitable
substrate for bioenergy application due to hydrophilic properties.
Adsorption of moisture may lead to a decrease in the energy den-
sity of the biomass [18]. Comparative low moisture content feed-
stock (<15%) is preferred when using a thermal conversion process,
while bio-conversion can utilize biomass with a high moisture con-
tent [19]. The moisture content of the biomass was determined us-
ing the procedure given in ASTM 3173-87 [20].
2.2.2. Ash content
Ash, the incombustible solid mineral matter present in the
biomass, mainly contains oxides of silica (SiO), Aluminium (AlO),
Iron (FeO), calcium (CaO) and Magnesium (MgO). The cations
present in ash retards the enzymatic saccharification of biomass
samples to glucose and other fermentable sugars [21]. Therefore it
is prerequisite to analyze the biomass for ash content before us-
ing it as a feedstock for biofuel production. Determination of ash
content in biomass samples were adopted as per the protocol de-
scribed in ASTM D 3174-04 [22]. Oven dried biomass sample (1.0 g)
was taken in oven dried moisture free crucibles and placed in muf-
fle furnace maintained at 575 ± 10 °C for 4 h. Crucibles were re-
moved from the furnace and placed in a dessicator and the pro-
cess of heating and cooling was repeated until constant weight was
obtained.
2.2.3. Volatile matter and fixed carbon
The volatile matter in the leaf litter biomass was determined
by the procedure given in ASTM D 3175-07 [23]. A biomass sam-
ple (1.0 g) was taken in a covered crucible and placed in a muf-
fle furnace regulated at 950 ± 10 °C for 7 min to obtain rapid heat-
ing. Then the crucible was removed from the furnace and placed
in a desiccator. The loss of weight was interpreted as the volatile
matter in the biomass. Fixed carbon was the resultant of the sum-
mation of percentage moisture, ash, and volatile matter subtracted
from 100.
2.2.4. Calorific value
The calorific value of all biomass samples was determined in
a static bomb calorimeter; BCM 211059, using the procedure de-
scribed in NREL protocol [24]. Each biomass sample was mixed
thoroughly in the sample bottle and was made into a pellet, tak-
ing care that the heavies and lights (fluff) were distributed well in
the sample. Samples (1.0 g) in the form of pellet was measured out
and put into pre-weighed crucibles. A cotton thread was attached
to a platinum ignition wire and placed in contact with the pellet.
Distilled water (1 ml) was poured into the bomb and then filled
with oxygen to a consistent pressure between 20 and 30 atm (2.03
and 3.04 MPa). The calorimeter was placed in an isothermal jacket
with an air gap separation of 10 mm between all surfaces. The
electrical energy for ignition, discharged through a platinum wire
of ∼40 V, was calculated using the change in potential across a
1256 or 2900 mF capacitor. The bomb calorimeter was submerged
in a calorimeter cane filled with distilled water. The jacket of the
calorimeter was maintained at constant temperature by circulating
water at 25° C.
2.2.5. Scanning electron microscopy (SEM)
The physical appearance of biomass samples were observed
by SEM. The samples were coated to provide conductivity with
gold using gold sputter at a voltage of 10–15 kV. Images were
taken using SEM (Model: JEOL JSM-6510 LV, USA) at different
magnifications.
2.2.6. X-ray diffraction (XRD) analysis
Crystallinity of the powdered biomass samples was analyzed at
room temperature in the scanning angle of 5–50° at the scan speed
of 5°/min using a PANalytical X’Pert PRO diffractometer (Nether-
lands) with Ni-filter, operated at 45 kV and 40 mA with λ (Cu
Kα ) = 1.5406 Å. The crystallinity index (CrI) of the biomass samples
were determined as described by Segal et al. [25] as follows:
CrI = [(I002 − Iam )/I002 ] × 100
Where I002 is the intensity for the crystalline portion of biomass
(i.e. cellulose) at about 2θ of 21–23°, and Iam is the peak for
the amorphous portion (i.e. cellulose, hemicellulose, and lignin) at
about 2θ of 16–18° in most literatures [1,2,17,26].
2.2.7. Thermal gravimetric analysis (TGA)
To understand the devolatilization characteristics, TGA of
biomass samples were performed using a Mettler Toledo instru-
ment (Model: TGA/SDTA 851e). The devolatilizations of biomasses
were studied from 20 to 700 °C with a heating rate of 10 °C/min
with a purge gas (nitrogen) flow rate of 60 ml/min.
2.3. Chemical characterization of biomasses
2.3.1. CHNSO analysis
The basic elements of any biomass such as carbon (C), hydro-
gen (H), nitrogen (N), sulphur (S) and oxygen (O), were analyzed
 N. Akhtar et al. / Journal of the Taiwan Institute of Chemical Engineers 62 (2016) 239–246 241

Fig. 1. Scheme for characterization of biomass samples.

Table 2
Proximate, ultimate and calorific value of biomass.

Biomass Proximate analysis (wt%) Ultimate analysis (wt%) Calorific Value

(MJ/kg)
§ ¥
Ash Volatile matter Fixed carbon Moisture C H N S O

M. indica 9.46 ± 0.60a 75.01 ± 4.77a 7.71 ± 0.40b 7.57 ± 0.47b 39.32 ± 2.40b 5.25 ± 0.20b 0.28 ± 0.01b 0.10 ± 0.008a 55.05 ± 3.95a 18.37 ± 0.46b
P. deltoides 3.70 ± 0.28c 76.05 ± 6.25a 11.23 ± 0.78a 8.32 ± 0.70a 40.68 ± 3.08b 4.81 ± 0.49c 1.40 ± 0.03a 0.02 ± 0.002b 53.09 ± 2.55a 18.50 ± 0.45b
P. longifolia 8.67 ± 0.65b 75.34 ± 4.18a 7.66 ± 0.71b 7.34 ± 0.69b 44.78 ± 2.39a 5.55 ± 0.15a 1.48 ± 0.09a 0.09 ± 0.004a 48.10 ± 2.50b 19.32 ± 0.73a

Values are mean ± SD (n = 3), bearing different letters in same column are significant at P < 0.05.
§
Percent of fixed carbon calculated from difference of moisture, ash and volatile matter content.
¥
Percent O calculated from the difference of C, H, N and S.

in a PerkinElmer Elementar CHNSO analyzer. The sample (1.0 mg)
was used in a tin boat assortment at 900 °C in an oxygen atmo-
sphere to determine the percentage composition of C, H, N and S.
The percentage of O was determined by means of difference.
2.3.2. Determination of extractives and compositional analysis
For determination of extractives, biomass samples (5 g) were
extracted using a Soxhlet extraction apparatus through cotton cel-
lulose extraction thimbles (external length 94 mm and inner diam-
eter 33 mm) as per the NREL protocol [24]. The heating mantle was
adjusted to provide a minimum of 6–10 siphon cycles per hour.
Three stages of extraction; n-hexane, ethanol and distilled water
were performed and each extraction operation was executed for
6 h. The solvent in each extract was removed in a rotary evapora-
tor under reduced pressure. In the first step the extraction of n-
hexane soluble compounds was carried out to separate the class
of compounds such as non-polar lipids, hydrocarbon compounds,
terpenoids etc. The raffinated biomass of the first step was ex-
tracted with ethanol to separate compounds such as chlorophyll,
polar waxes, sterol and other minor constituents. The final raffinate
biomass was extracted with distilled water to separate inorganic
materials and non-structural carbohydrates [1]. Raffinate biomass
was subjected to cellulose, hemicelluloses and lignin estimation by
TAPPI protocol [27].
2.3.3. Fourier transform infrared (FTIR) spectroscopy
Accurate predictions of the chemical composition of biomass
samples were obtained by FTIR spectroscopy using a Nicolet Instru-
ment (Model: MAGNA 550, USA). The biomass sample (10 mg) was
mixed with 200 mg of KBr and the mixture was compressed for
preparation of pellets. Each spectrum was the average of 64 scans
with a total scan time of 15 s in the IR range of 40 0–40 0 0 cm−1 .
2.3.4. Solid state 13 C CP/MAS NMR spectroscopy
Solid state 13 C CP/MAS NMR spectroscopy is an important tool
to analyze the structural features of lignocellulosic biomass. The
spectra of biomass samples were obtained at 75.475 MHz using
a Bruker AV-300 FT NMR spectrometer (Germany). Dried samples
were spun at a rate of 10 kHz using 4 mm zirconia (ZnO2 ) rotor
with CP pulse programme. Typically 18,0 0 0 transients were accu-
mulated in the time domain with a contact time of 1 ms and a re-
cycle delay of 3 s. Tetramethylsilane (TMS) was used as an external
standard for the calibration of the 13 C chemical shift.
2.3.5. Statistical analysis
All the experiments were performed in triplicate and the data
were analyzed by analysis of variance (ANOVA) using GraphPad
Prism (5.0) software followed by the Tukey’s honestly significant
difference test. Results were expressed as mean ± S.D. (n = 3).
3. Results and discussion
3.1. Determination of proximate, ultimate and calorific values
The proximate and ultimate composition along with calorific
value of leaf litter biomasses from Mango, Poplar and Ashoka are
shown in Table 2. The percentage of ash in Mango, Poplar and
Ashoka leaf litters were found to be 9.46, 3.70 and 8.67%, respec-
tively. Moisture content of these biomass samples were found in
the range of 7.34–8.32%. Similar results were observed for wheat
straw, barley straw, flax straw and timothy grass and reported
in the range of 5.0–7.9% for moisture and 1.1–9.8% for ash con-
tent by Naik et al. [1]. Biomass of Areca catechu, Ziziphus rugosa
and Albizia lucida showed moisture content of 6.6, 5.2 and 5.8%
and ash content of 2.9, 1.3 and 1.5%, respectively [17]. Greater
amount of ash was observed with Mango and Ashoka leaf lit-
ter biomasses, accounted them less suitable than Poplar leaf litter
biomass for bioethanol production as ash cannot be converted into
energy. Feedstock with low moisture content (<15%) is preferred
using thermal conversion process, while conversion using biolog-
ical means require relatively high moisture for submerged and
solid state fermentation. In biomass fuel, chemical energy is stored
mainly in the form of fixed carbon and volatile matter, which can
be released via direct or indirect combustion. High volatile matter
and low ash content were observed with Poplar leaf litter biomass,
may be considered as promising substrate for biofuel production.
242 N. Akhtar et al. / Journal of the Taiwan Institute of Chemical Engineers 62 (2016) 239–246

Table 3
Peak at crystalline and amorphous region and CrI of different biomass samples.

Biomass Crystalline portion peak Amorphous portion peak CrI (%)

at 2 θ angle at 2 θ angle

M. indica 22.89° 18.45° 23.13

P. deltoides 22.82° 18.62° 20.93
P. longifolia 22.30° 18.49° 20.94

CHNSO analysis of biomass samples showed no marked differ-

ence (Table 2). Leaf litter biomass samples contained C, H, N, S and
O in the range of 39.32–44.78%, 4.81–5.55%, 0.28–1.48%, 0.02–0.10%
and 48.10–55.05%, respectively. Results of ultimate analysis were
in good agreement with the earlier results reported by Naik et al.
[1] and Sasmal et al. [17] with different biomass samples. Nitrogen
and S generates oxides as gaseous emissions (NOx, SOx) on com-
bustion, therefore does not contribute to the total heating value
of biomass. In contrast to conventional fuels, biofuels demonstrate
relatively low proportions of C, as compared to the total proportion
of O and H, deteriorating fuel energy value due to lower energy
contained in C =O and C–H bonds over C–C bonds [28]. Calorific
value of Ashoka leaf litter biomass was found greater (19.32 MJ/kg)
as compared to Poplar (18.50 MJ/kg) and Mango (18.37 MJ/kg) leaf
litter biomasses. High calorific value of wheat straw (20.3 MJ/kg),
pinewood (19.6 MJ/kg) [1], Z. rugosa (21.24 MJ/kg) and A. lucida
(20.35 MJ/kg) [17] makes these biomass types more efficient for
production of bio-energy than the litters we studied.

3.2. Scanning electron microscopy (SEM)

Scanning electron micrographs of native biomass fractions ex-

hibited different surface structure with intact morphology. Distinct
surface structures were observed with micrographs (Fig. 2) of dif-
ferent leaf litter biomasses from Mango, Poplar and Ashoka, prob-
ably due to taxonomic differences. More compact structure was
seen with the micrograph of Mango leaf litter (Fig. 2a). However,
comparatively less compact and fibrillar structure was observed
with leaf litter biomasses of Poplar (Fig. 2b) and Ashoka (Fig. 2c).
Rigid and compact structure is generally observed due to encap-
sulation of cellulosic fibers by lignin-hemicellulose matrix, which
hinders accessibility of cellulose to hydrolytic enzymes for conver-
sion processes.

3.3. X-ray diffraction (XRD) analysis

The XRD analysis of leaf litter biomass samples are presented in

Fig. 3. The peaks for crystalline and amorphous cellulose were ob-
served near 2θ angles of 18° and 22°, respectively (Table 3, Fig. 3)
for leaf litter biomasses from Mango, Poplar and Ashoka. CrI of
these samples was found in the range of 21–23% in the native
state. High crystallinity hinders enzymatic hydrolysis of lignocel-
lulosic biomass. Amorphous cellulose is readily accessed by cellu-
lases as compared to crystalline microfibrils [29] and it is widely
accepted that the decrease in crystallinity increases the digestibil-
ity of lignocelluloses [30,31]. On the contrary, more crystalline lig-
nocelluloses have been reported to show more digestibility [2,32].
Similar peaks for crystalline and amorphous cellulose were also
observed with different biomass samples [1,17,26,32]. Crystallinity
depends on relative amounts of crystalline and amorphous compo-
nent of cellulosic microfibrils [29]. However, determining true crys-
tallinity of cellulose is ambiguous, as crystallinity depends upon
the entire material including hemicelluloses, lignin and the nature
of bonds between them too. Therefore, further study between ma-
jor chemical classes and their interactions is necessary to justify
the diffractographic studies.
Fig. 2. Scanning electron micrograph of leaf litter biomass samples from (a) Mango,
(b) Poplar and (c) Ashoka (Magnification: 1,0 0 0X).
3.4. Thermal gravimetric analysis (TGA)
The behavior of different biomass samples during devolatiliza-
tion were quite different (Fig. 4.). The onset temperature of de-
volatilization was in the range of 200–250 °C, which corresponded
to weight loss (∼10%) with respect to the final weight loss. The
main weight loss ends at 310–350 °C for all biomasses and then
followed by a slow and continuous weight loss. The former step
was due to the primary devolatilization, whereas the later was
attributed to the degradation of heavier chemical structures in
the solid matrix [33]. The weight loss during devolatilization can
be explained as follows: the weight loss at temperature less
than 100 °C was for loss of easily volatilized compounds, 100–
130 °C for loss of water, 130–250 °C for loss of volatile compounds,
 N. Akhtar et al. / Journal of the Taiwan Institute of Chemical Engineers 62 (2016) 239–246 243

Table 4
Extractives (g) from 100 g of biomass samples.

Biomass Hexane extract Ethanol extract Water extract Cellulose Hemicellulose Lignin

M. indica 9.65 ± 0.55b 8.68 ± 0.55b 13.39 ± 0.92a 31.23 ± 1.71b 28.02 ± 3.02b 10.56 ± 0.79b
P. deltoides 8.60 ± 0.54c 9.79 ± 0.52a 9.95 ± 0.73b 37.75 ± 4.21a 21.85 ± 1.55c 11.95 ± 0.80a
P. longifolia 10.59 ± 0.81a 10.26 ± 0.66a 8.84 ± 0.53c 26.65 ± 1.64c 32.04 ± 2.19a 10.63 ± 1.18b

Values are mean ± SD (n = 3), bearing different letters in same column are significant at P < 0.05.

litter biomasses may be attributed to high mannose or lignin con-

tent [1]. Similar devolatization patterns were observed with differ-
ent feedstocks such as rice straw, wheat straw, barley straw, flax
straw, pine wood [1], Tamarix ramosissima [2], A. catechu, A. lucida
and Z. rugosa [17], characterized for biofuel production.

3.5. Biomass extractives

The yield of hexane, ethanol and water extracts along with cel-
lulose, hemicelluloses and lignin content of different biomass sam-
ples are presented in Table 4. In terms of total extract, leaf lit-
ter biomass of Mango has maximum extractives (31.72 wt%) fol-
lowed by Ashoka (29.69 wt%) and Poplar (28.34 wt%). Among three
leaf litter biomass studied, Poplar showed greater cellulose con-
tent (37.75 wt%) followed by Mango (31.23 wt%) and Ashoka (26.65
wt%), however, hemicellulose content was in the range of 22–32%.
Comparable range of cellulose (29–39%) and hemicelllulose (26–
34%) content was observed in wheat straw, barley straw, flax straw,
trimothy straw and pinewood [1]. However, greater cellulose (45–
53%) with less hemicelllulose (6–28%) content was observed with
the biomasses of A. catechu, A. lucida, Z. rugosa [17]. Cellulosic and
hemicellulosic component of these biomasses could be used as po-
tential substrate for enzymatic hydrolysis to obtain fermentable
Fig. 3. X-ray diffractograph of leaf litter biomass samples from (a) Mango, (b)
sugar for bioethanol production.
Poplar and (c) Ashoka.

3.6. Fourier transform infrared (FTIR) spectroscopy

Fourier transform infrared (FTIR) spectroscopy analysis was car-

Fig. 4. Thermogravimetric analysis of leaf litter biomass samples from (a) Mango,
(b) Poplar and (c) Ashoka.
250–350 °C for loss of hemicellulose, 350–500 °C for loss of cel-
lulose, lignin and > 500 °C for lignin. A sharp decline beyond
500 °C was observed with Mango leaf litter biomass, which gener-
ally shows more cellulose and hemicelluloses content, whereas the
slow rate of devolatilization pattern with Ashoka and Poplar leaf
ried out to detect changes in functional groups of different biomass
samples. The obtain FTIR spectrum is represented in Fig. 5 and
assigned fingerprints are summarised in Table 5 based on litera-
ture values [1,2,17,34,35]. The main characteristics were attributed
to different chemical groups present in cellulose, hemicellulose
and lignin components of biomasses. The greater percent trans-
mittance was observed for leaf litter biomass of Poplar followed
by Ashoka and Mango in the range 360 0–30 0 0 cm−1 . This range
represents the hydrogen bonded stretching bands of OH groups, ei-
ther from the glucoside linkages of cellulose or the hydroxyphenyl,
guaiacyl and syringyl groups of lignin [36]. Lignocellulosic biomass
most likely consisted of alkene, esters, aromatics, ketone and alco-
hol, with different oxygen-containing functional groups. The wave
number 140 0–90 0 cm−1 is designated to vibrations of a variety
of bonds such as C–H, C–O, C–N and P–O [37]. Absorption for
wave number in between 1450 and 10 0 0 cm−1 corresponds to a
large family of esters formed between quinic acid and one to four
residues of certain trans-cinnamic acids, most commonly caffeic,
p-coumaric and ferulic [38], contributing to majority of observed
bands. The readily visible peak at 1635 cm−1 was observed in all
three types of biomass samples, and is generally attributed to C=O
stretching of hemicelluloses and deformation vibrations of H–O–H
in absorbed water [2]. The prominent peak near 1043 cm−1 cor-
responded to C–O stretching vibration in cellulose and hemicellu-
loses, advocated that all leaf litter biomass samples are rich source
of cellulose and hemicellulose and can be used as a promising
feedstock in biorefinery.
244 N. Akhtar et al. / Journal of the Taiwan Institute of Chemical Engineers 62 (2016) 239–246

Table 5
Assignment of FTIR bands of functional groups in different leaf litter biomass.

Name of characteristic group Wavenumber (cm−1 )

O–H stretching in cellulose and lignin 360 0–30 0 0

C–H stretching of aliphatic structure 2960–2850
C=O stretching of hemicelluloses 1750–1630
C–H deformation in cellulose and hemicellulose 1384–1346
Syringyl ring breathing and C–O stretching out of lignin and xylan ∼1232
C–O stretching vibration in cellulose and hemicelluloses ∼1043

Table 6
13
Signal assignments for chemical shifts obtained from solid state C CP/MAS NMR spec-
troscopy of different leaf litter biomass.

Chemical shift (ppm) Assignments

173 Carboxyl group of hemicelluloses

104 Anomeric carbon (C1) of cellulose
83 C4 of amorphous cellulose and hemicelluloses
72 C2, C3, C5 of cellulose, OCα H2 carbon of lignin
63 C6 of crystalline cellulose
32 Methoxyl (–OCH3 ) and methylene (–CH2 ) groups in lignin
23 CH3 in acetyl group of hemicelluloses

Fig. 6. Solid state 13 C CP/MAS NMR spectra of leaf litter biomass samples from (a)
Mango, (b) Poplar and (c) Ashoka.
Fig. 5. FTIR spectra of leaf litter biomass samples from (a) Mango, (b) Poplar and
(c) Ashoka.

content of hemicellulose. The conclusions drawn from NMR spec-

3.7. Solid state 13 C CP/MAS NMR spectroscopy
Solid state 13 C CP/MAS NMR spectra provide characteristic
chemical shift values ascribed to cellulose, hemicellulose and
lignin, represented in Fig. 6. Signal assignments for various
biomass samples have been assigned based on the literature
[39–41] and summarised in Table 6. The prominent peak at
72 ppm along with peaks at 104, 83 and 63 ppm render a clue for
the presence of carbohydrates in Mango, Poplar and Ashoka leaf
litter biomass samples. A prominent peak at 23 ppm was typically
observed with biomass samples of Ashoka, advocating for a greater
tra (Fig. 5) support the result obtained from chemical analysis
of biomasses for cellulose and hemicellulose content (Table 4).
Presence of cellulose and hemicelluloses content of these biomass
samples recommend their use as feedstock for bio-energy gen-
eration. Intense peaks assigned in the region of 50–120 ppm are
attributed mostly to cellulose carbon, contributions from hemicel-
lulose along with some lignin signals. However, signals from lignin
are generally concentrated between 10 0 and 20 0 ppm. An aliphatic
region for lignin lies from 0 to 95 ppm whereas, an aromatic region
falls in between 100 and 160 ppm [42]. Chemical shift deflection
for C4 and C6 carbon in the anhydroglucose unit rely upon the cel-
 N. Akhtar et al. / Journal of the Taiwan Institute of Chemical Engineers 62 (2016) 239–246
lulosic source, variations in cellulose crystallinity and crystal lattice
structure [43].
4. Conclusion
Proper utilization of forest and agricultural residues is a healthy
symbol for an economically viable and environment friendly com-
munity. Physico-chemical characterization of leaf litter biomasses
from different trees such as Mango, Poplar and Ashoka suggested
that it could be used as feedstock for gasification, bio-oil and bio-
alcohol production and to meet the demand of second genera-
tion biofuel. To the best of our knowledge, this is the first sys-
tematic report on critical evaluation of leaf litter biomass based
on physico-chemical characteristics for their use as feedstock in
bio-energy production. The ideal characteristics of biomass sam-
ples for production of biofuel are greater calorific value along with
greater cellulose and hemicelluloses content as compared to ash
and lignin. However, selection of suitable feedstock for bio-energy
production based on mere physico-chemical characteristic is not a
wise decision, several factors such as crystallinity, degree of poly-
merization, particle size and accessible surface area of biomass
often limits conversion process. Delineation of physico-chemical
characteristics of these biomasses surely proves their potential over
present day conventional substrates such as sugarcane bagasse and
corn. The compositional analysis needs special care and consider-
ation, including sample drying, size reduction, presence of non-
structural carbohydrates, extractable ash, proteins, starch and high
extractive contents [44]. FTIR is the simplest method which re-
quires very less amount of sample without the necessity of specific
operation experience; however, it gives only relative values not the
absolute values, as contributions from both crystalline and amor-
phous regions are present in the FTIR spectrum [45]. In addition
to accuracy in measurement of crystallinity, solid state 13 C NMR
spectroscopy provides detailed structural elucidation of the ligno-
cellulosic components [46]. Optimal utilization of feedstock for bio-
fuel generation requires efficient pretreatment techniques, robust
techno-economic analysis such as energy balance, low doses of en-
zyme loading, enhanced saccharification and solvent recovery for
their successful implementation in biochemical refineries.
Acknowledgments
The authors are thankful to Directors, Thapar University and IIT
Guwahati for providing infrastructural support. The research work
by A. Goyal and D. Goyal was supported by joint project grant from
Department of Biotechnology, Ministry of Science and Technology,
Government of India (BT/23/NE/TBP/2010).
Supplementary materials
Supplementary material associated with this article can be
found, in the online version, at doi:10.1016/j.jtice.2016.02.011.
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