Journal of Macromolecular Science, Part A

Pure and Applied Chemistry

ISSN: 1060-1325 (Print) 1520-5738 (Online) Journal homepage: https://www.tandfonline.com/loi/lmsa20

Formulation and characterization of in situ

generated copper nanoparticles reinforced
cellulose composite films for potential
antimicrobial applications

P. Sivaranjana, E. R. Nagarajan, N. Rajini, M. Jawaid & A. Varada Rajulu

To cite this article: P. Sivaranjana, E. R. Nagarajan, N. Rajini, M. Jawaid & A. Varada Rajulu
(2018) Formulation and characterization of in situ generated copper nanoparticles reinforced
cellulose composite films for potential antimicrobial applications, Journal of Macromolecular
Science, Part A, 55:1, 58-65, DOI: 10.1080/10601325.2017.1387488

To link to this article: https://doi.org/10.1080/10601325.2017.1387488

Published online: 01 Dec 2017.

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JOURNAL OF MACROMOLECULAR SCIENCE, PART A: PURE AND APPLIED CHEMISTRY
2018, VOL. 55, NO. 1, 58–65
https://doi.org/10.1080/10601325.2017.1387488

Formulation and characterization of in situ generated copper nanoparticles reinforced

cellulose composite films for potential antimicrobial applications
P. Sivaranjanaa, E. R. Nagarajana, N. Rajinib, M. Jawaidc, and A. Varada Rajulud
a
Department of Chemistry, Kalasalingam University, Krishnankoil, Tamil Nadu, India; bCentre for Composite Materials, Department of Mechanical
Engineering, Kalasalingam University, Krishnankoil, Tamil Nadu, India; cLaboratory of Biocomposite Technology, Institute of Tropical Forestry and Forest
Products (INTROP), Universiti Putra Malaysia, Serdang, Selangor, Malaysia; dCentre for Composite Materials, International Research Centre, Kalasalingam
University, Krishnankoil, Tamil Nadu, India

ABSTRACT ARTICLE HISTORY

Cellulose was dissolved in aq.(LiOH C urea) solution pre-cooled to –12.5 C and the wet films were Received March 2017
prepared using ethyl alcohol coagulation bath. The gel cellulose films were dipped in 10 wt.% Cassia alata Revised July 2017
leaf extract solution and allowed the extract to diffuse into them. The leaf extract infused wet cellulose Accepted September 2017
films were dipped in different concentrated aq. copper sulphate solutions and allowed for in situ KEYWORDS
generation of copper nanoparticles (CuNPs) inside the matrix. The morphological, structural, antibacterial, Green composites; copper
thermal, and tensile properties of dried cellulose/CuNP composite films were carried out. The presence of nanoparticles; in situ
CuNPs was established by EDX spectra and X-ray diffraction. The composite films displayed higher thermal generation; nano-
stability than the matrix due to the presence of CuNPs. Cellulose/CuNP composite films possessed better composites; antibacterial
tensile strength than the matrix. The composite films showed good antibacterial activity against E.coli activity; tensile strength
bacteria. We conclude that good antibacterial activity and better tensile properties of the cellulose/CuNP
composite films make them suitable for antibacterial wrapping and medical purposes.

1. Introduction
The synthetic polymers and their composites though find innu-
merable applications in our daily life, do not degrade and hence
pose many environmental problems. Further, the synthetic pol-
ymers are derived from precious petroleum which is fast
depleting. Hence, there is a need to look at safer alternatives for
synthetic polymers. In recent years, many researchers are work-
ing to develop biodegradable polymers and their composites
from renewable sources. In this direction, Feldman[1] prepared
lignin nanocomposites using nanolignin as filler and lignin as
matrix. Li et al.[2] prepared polypropylene carbonate/Hildegar-
dia short fiber completely biodegradable composite films for
packaging applications. Chen et al.[3] prepared fully biodegrad-
able poly (lactic acid)/poly (propylene carbonate)/poly (buty-
lenes adipate-co-terephthalate) blends combined with
chemometrics and characterized them. Jayaramudu et al.[4]
used natural fibers of Sterculia urens in polylactic acid matrix
to prepare completely biodegradable polymer composites and
recommended them for packaging applications. Recently, Feng
et al.[5] and Xia et al.[6] used egg shell powder and spent tea leaf
powder respectively as fillers in polypropylene matrix and pre-
pared green composites with improved properties. Liu and
Zhou[7] synthesized biodegradable poly (butylene succinate-co-
butylene itaconate) copolymers and characterized them. Simi-
larly, Jagadeesh et al.[8] prepared wheat protein isolate/Hilde-
gardia completely biodegradable composites for packaging
applications. Jeevan Prasad Reddy et al.[9] prepared soy protein
isolate films for packaging photosensitive materials. But of all
the biodegradable matrices, cellulose is the most abundant
nature’s gift which can be converted into films or can be modi-
fied to make useful articles. Some researchers used curcu-
min,[10] Sterculia urens,[11] spent tea leaf powder,[12] waste silk
fiber[13] etc. as fillers in cellulose matrix and prepared
completely biodegradable composite films.
For medical applications, the composites should possess
good antibacterial activity. Recently, Feldman[14] in his review
article discussed the importance of polymer nanocomposites in
medicine. Feldman[15] also reviewed the preparation, character-
ization and applications of polyblend nanocomposites. As
metal nanoparticles possess good antibacterial activity, some
composites were made using them as the fillers. Recently,
Sadanand et al.[16] prepared cellulose/silver nanoparticle
(AgNP) nanocomposite films with in situ generated AgNPs
using Ocimum sanctum leaf extract as a reducing agent. Sada-
nand et al.[17] also prepared cellulose/copper nanoparticle
(CuNP) composite films using the same Ocimum sanctum leaf
extract as a reducing agent. The properties of the in situ gener-
ated metal nanoparticle composites by green synthesis depend
on the nature of the leaf extract used for reducing copper salts
into CuNPs. In the present work, the authors used the antibac-
terial Cassia alata leaf extract as a reducing agent and aq.
CuSO4.5H2O as a source solution for in situ generation of
CuNPs. The authors selected Cassia alata leaf extract in
the present work as it possesses many medicinal properties.

CONTACT N. Rajini rajiniklu@gmail.com Department of Mechanical Engineering, Kalasalingam University, Krishnankoil – 626126, Tamil Nadu India.
Color versions of one or more of the figures in the article can be found online at www.tandfonline.com/lmsa.
© 2018 Taylor & Francis Group, LLC
 JOURNAL OF MACROMOLECULAR SCIENCE, PART A: PURE AND APPLIED CHEMISTRY
Paul et al.[18] isolated an antibacterial constituent from the
leaves of Cassia alata Linn and reported that the main compo-
nent in this leaf – 3,4 dihydroxycinnamic acid had antibacterial
activity against both Gram-positive and Gram-negative bacte-
ria. Faruq et al.[19] extracted the chemical components of Cassia
alata leaves in different solvents and came to the conclusion
that only one component mainly composed of steroids was
responsible for the maximum antibacterial activity.
In the present work, in addition to the in situ generation of
CuNPs in the cellulose matrix, some CuNPs were also formed
outside the composite films in the copper sulphate solutions
used for the generation of CuNPs. The CuNPs formed outside
the composite films were isolated, cleaned and observed under
Transmission electron microscope (TEM) and scanning elec-
tron microscope (SEM) and the particle size distribution was
examined. The cellulose/CuNP composite films were subjected
to SEM. Fourier transform infrared (FTIR) spectroscopy, X-ray
diffraction (XRD), Thermogravimetric Analysis (TGA), anti-
bacterial activity and tensile tests.
2. Materials and method
2.1. Materials
In the present work, cotton linters (with a degree of polymeri-
zation Dp of 620) supplied by Hubei Chemical Fiber Co. Ltd.
(Xiangfan, China) was used. LiOH, CuSO4.5H2O and Urea
supplied by Hi Media laboratories Pvt. Ltd (India) were used
without further purification. Ethyl alcohol was supplied by
Ganesh Scientific (India) suppliers. The Cassia alata leaves
were collected locally and washed thoroughly with distilled
water before preparing the extract.
2.2. Cassia alata leaf extraction
The Cassia alata leaves were collected, washed and cut into
small pieces of even size. 10 wt% of leaves were taken in dis-
tilled water boiled over water bath maintained at 80 C for
about 20 minutes. Then the solution was cooled, filtered and
the supernatant liquid was stored.
2.3. Dissolution of cellulose
Cellulose solution was made as described earlier.[16] In brief,
the aqueous solution of 8 wt% of LiOH and 15 wt% of urea
was prepared and refrigerated to –12.5 C. 4 wt % of cotton
linters were added to the solution and stirred using a
mechanical stirrer and within 2 minutes the cellulose
(cotton linters) got dissolved completely. The residues
remaining undissolved were removed by centrifugation at a
speed of 7200 rpm and temperature of 5 C. The clear stock
solution was stored at 5 C till used.
2.4. Preparation of cellulose wet films with diffused leaf
extract
The cellulose solution was spread on glass plates using spacers
and regenerated in alcohol bath. The wet cellulose films were
thoroughly washed in distilled water several times for the
59
removal of alcohols and dissolved salts if any. These wet films
were kept inside the leaf extract for 24 hrs for its diffusion into
the films.
2.5. Preparation of cellulose/CuNP composite films
Copper sulphate solutions of varying concentrations of 5 mM,
25 mM, 125 mM and 250 mM were prepared. Each concen-
trated solution was kept in separate beakers and the films
infused with leaf extracts were immersed in them separately.
Soon the color of the wet matrix films started changing indicat-
ing the reduction of copper sulphate to CuNPs. After 24 hrs,
the films were taken out and washed thoroughly with distilled
water. The color of the films was retained even after the
repeated washings which indicated that stable nano particles
were formed inside film. The films were then dried over a
smooth surface at room temperature.
2.6. Characterization
To examine the color formation of the cellulose/CuNP compos-
ite films, the UV-visible spectra of the matrix and the nano-
composite films were recorded using UV-–1800 SHIMADZU
spectrophotometer. The FTIR spectra of the leaf extract infused
cellulose (matrix) and the composite films were recorded on a
spectrum RXI Perkin Elmer FTIR spectrophotometer. All the
spectra were recorded in each case only once from 4000 pcm¡1
to 500 cm¡1 range with 45 scans at a resolution of 4 cm¡1. The
morphology of the films and the distribution of CuNPs inside
the films were examined with Carl Zeiss EVO 18, scanning
electron microscope and the EDX spectra were also recorded
with BRUKER X FLASH6130 detector attached to the micro-
scope using ESPIRIT software. The nano particles formed out-
side the films in solution were examined for their particle size
distribution with Joel model JEM 210 transmission electron
microscope and SEM. X-Ray diffractograms of the matrix and
cellulose/CuNP composite films were recorded using a Bruker
Eco D8 Diffractometer in the 2u range of 10 to 90 operated at
a voltage of 40 kV and current 25 mA.
The primary and derivative thermograms of matrix and cel-
lulose/CuNP composite films were recorded using TGA Q500
TA Instruments, USA in the temperature range of 50 – 600 C
in nitrogen atmosphere at a heating rate of 10  C/min. The ten-
sile strength of the matrix and the nanocomposite films was
determined using 5 kN INSRON tensile machine with an initial
grip separation and crosshead speed of 50 mm and 5 mm/min,
respectively. Filmstrips cut into a length of 100 mm and width
10 mm were used for this work. For each category, 5 specimens
were tested and the average values are reported.
2.7. Antibacterial test
The antibacterial test was conducted as described in our previ-
ous work.[16,20] In brief, 200 ml of LB and agar containing
medium was prepared and sterilized for 20 minutes and the
prepared medium was immediately transferred into sterile Petri
plates. The plate was left for 15 minutes to solidify. After solidi-
fication, overnight cultured E.coli was taken on sterile buds and
was swapped on the surface of the agar plates. Wells were
60 P. SIVARANJANA ET AL.

punched on each plate using gel puncture. The circular discs of

the matrix and the cellulose/CuNP composite films with
CuNPs generated using 5 mM, 25 mM, 125 mM and 250 mM
aq.CuSO4.5H2O solutions were tested against pathogenic E.Coli
bacteria. The Zones of clearance formed were recorded and
their diameters were measured. We selected the disk method,
as it is simple and conventionally accepted.

3. Results and discussion

3.1. Appearance of matrix and cellulose/CuNP composite
film
In order to examine the appearance of the matrix and cellulose/
CuNP composite films, their photographs were recorded and
are presented in Figure 1. From Figure 1, it is evident that the
matrix was light brown in color while the color of composite
films changed from medium brown to dark brown with
increasing concentration of copper sulphate solutions. This
indicates the in situ generation of CuNPs inside the matrix. In
addition to the visual observation, the UV-Visible spectra of
the matrix and the cellulose/CuNP composite films with in situ
generated CuNPs using different concentrated source solutions
were recorded to confirm the differences in the optical appear-
ance between the matrix and the nanocomposite films. The
UV-Visible spectra are presented in Figure 2. From Figure 2, it
can be seen that the nanocomposite films exhibited higher
absorbance than the matrix film and increased with increasing
concentration of the source solutions. Further, the maximum
absorbance for the matrix appeared at 386 nm while for the
nanocomposite films made with higher concentrated source
solutions, it shifted to 404 nm. These differences indicate the in
situ generation of CuNPs in the composites.
3.2. Particle size distribution of CuNPs formed outside the
matrix
In addition to the in situ generation of CuNPs in the matrix,
some nano particles were also formed outside the matrix film
(due to the interaction of some diffused extract from the matrix
into source solutions) which were separated by successive cen-
trifugations and dried. To examine the nature of CuNPs
formed outside the matrix, their TEM and SEM images were
recorded which are presented in Figure 3a and Figure 3b
respectively. The EDX spectrum of the CuNPs is presented in
Figure 3c. Using the SEM micrograph, the particle size of one
Figure 2. UV absorbance spectra of matrix and cellulose/CuNP composites formed
using 5 mM, 25 mM, 125 mM and 250 mM aq. CuSO4.5H2O solutions
hundred CuNPs was measured and the particle size distribu-
tion is presented in Figure 3d. From Figure 3 (a, b), it is evident
that the CuNPs formed outside the matrix film were spherical
in shape and the average size of one hundred particles selected
was »75 nm. The presence of CuNPs was confirmed by EDX
spectrum. Further most of the CuNPs produced had the size
distribution in the 60–69 nm range.
3.3. Particle size distribution of in situ-generated CuNPs
inside the matrix
In order to confirm the in situ generation of CuNPs in the
matrix and to examine the particle distribution, the fractro-
graphs of cellulose/CuNP composite films were recorded. As
an example, the fractrographs of the composite films prepared
using 5 mM and 250 mM aqueous copper sulphate solutions
are presented in Figure 4a and Figure 4b respectively. The cor-
responding EDX spectra and the particle size distribution are
also presented in Figure 4(c, d). From Figure 4(a, b), it is evi-
dent that the in situ generated CuNPs in the matrix were also
spherical in shape. The formation of CuNPs was confirmed by
the corresponding EDX spectra. In each case, one hundred par-
ticles were selected randomly and their average particle size
was measured. The particle size distribution curves of in situ
generated CuNPs in the composites prepared using 5 mM and
250 mM aqueous copper sulphate solutions are presented in
Figure. 4e and Figure 4f respectively. The average particle size

Figure 1. Photographs of films of (a) cellulose with leaf extract (matrix), (b, c, d, and e) composites with CuNPs generated using aq.CuSO4.5H2O solutions of concentra-
tions 5 mM, 25 mM, 125 mM and 250 mM respectively.
 JOURNAL OF MACROMOLECULAR SCIENCE, PART A: PURE AND APPLIED CHEMISTRY 61

Figure 3. (a)Transmission electron micrograph (b) Scanning electron micrograph of CuNPs generated outside the film in solution. (c) EDX spectra and (d) Particle size dis-
tribution of CuNPs generated outside the film in solution.

in the case of the composites prepared using 5 mM and
250 mM aqueous copper sulphate solutions was found to be
49 nm and 70 nm respectively. The increased particle size
when higher concentrated CuSO4.5H2O solution was used indi-
cates some agglomeration. Further in the case of nano compo-
sites prepared using 5 mM aqueous copper sulphate solution,
most of the CuNPs were in the size range of 40–49 nm
(Figure 4e) whereas when 250 mM CuSO4.5H2O solution was
used, the CuNPs had the particle size distribution in the 70–
79 nm range (Figure 4f).
3.4. Interaction between matrix and CuNPs
In order to probe the interactions between matrix and CuNPs,
the FTIR spectra of the matrix and the composites were
recorded, and are presented in Figure 5. From Figure 5, it is evi-
dent that the spectrum of matrix is similar to that of the com-
posites. It clearly indicates that the interaction between the
matrix and CuNPs was mainly due to electrostatic forces.
The common bands observed at 3320, 2890, 1425, 1370,
1260 and 1158 cm¡1 belong to cellulose-II structure.[17]
Another prominent peak observed at 1644 cm¡1 was due to
crystallization of water.[16] The band at 900 cm¡1 was due to
C-OH groups of cellulose. As Cassia alata leaf extract has 3,
4-dihydroxycinnamic acid as the active compound,[18] the
peaks corresponding to -OH groups of the extract also over-
lap with those of cellulose. Further, the band at 931cm-1 was
attributed to OH bending of carboxylic acids present in the
leaf extract. Similarly, the peaks in the 900 to 666 cm¡1
region were due to -CH of aromatics of leaf extract. As the
baseline was not the same in the spectra of matrix and the
nanocomposite films, we could not make quantitative com-
parison of intensities.
3.5. Antibacterial activity
To probe the antibacterial activity of the composites, the anti-
bacterial activity test was conducted against E.coli bacteria by
disk method. As the leaves of Cassia alata also have antibacte-
rial activity, for comparison, the test for the matrix was also
conducted separately. The clear zones observed for the matrix
and the composites were photographed and are presented in
Figure 6a and Figure 6b respectively. In each case, the diameter
of clear zone was measured and the values are presented in
Table 1. From Figure 6a, it is evident that though the leaves of
Cassia alata possess antibacterial activity, the matrix exhibited
only a marginal activity. On the other hand from Figure 6b, it
can be seen that the composite films showed good antibacterial
activity against E.coli pathogen. From Table 1 it can also be
observed that the clear zone diameter increased with increasing
concentration of aqueous CuSO4.5H2O solutions used as the
source for the generation of CuNPs. The antibacterial activity
of the present system is compared with that of other similar
systems reported in the literature and is presented in Table 1.
3.6. X-Ray diffraction analysis
Inorder to study the effect of CuNPs on the crystalinity of the
composites, their X-ray diffractograms were recorded. The dif-
fractograms of the matrix and cellulose/CuNP composite films
formed using different concentrated copper sulphate solutions
are presented in Figure 7a. From Figure 7a, it is evident that there
are two main peaks in the case of matrix and the composites at
2u D 12.8 and 21.9 corresponding to reflections from (110)
and (200) respectively[21] but in the case of the composites the
peak at 21.9 was resolved into an additional peak at 19.8 that
corresponds to (110) plane of cellulose. Furthur there are two
62 P. SIVARANJANA ET AL.

Figure 4. (a) and (b) Scanning electron micrographs of CuNPs formed inside the composite films using 5 mM and 250 mM aq. CuSO4.5H2O solutions respectively. (c) and
(d) EDX spectra of CuNPs formed inside the composite films using 5 mM and aq. CuSO4.5H2O solutions respectively, (e) and (f) particle size distribution of CuNPs formed
inside the composite films using 5 mM and 250 mM aq. CuSO4.5H2O solutions respectively.

Figure 5. FTIR spectra of matrix and cellulose/CuNPcomposites formed using
5 mM, 25 mM, 125 mM and 250 mM aq. CuSO4.5H2O solutions.
more broad peaks at around 32.9 and 40.2 which may be due
to diffraction from the compounds of the leaf extract. On close
examination of the diffractograms corresponding to the compo-
sites, some additional peaks were observed (shown separately for
the composite with CuNPs formed using 250 mM copper sul-
phate solution by highlighting the 25 to 80 region in Figure 7b).
From Figure 7b some peaks were identified at 2u D 32.3 , 36.8 ,
43.6 and 51 . Of these, the peaks at 32.3 and 36.8 were
assigned to Cu2O nanoparticles with diffractions from (002) and
(111) planes respectively wereas the other two peaks at 43.6 and
51 were assigned to Cu nanoparticles with diffractions from
(111) and (200) planes respectively. Similar observation was
made by Sadanand et al.[17] in the case of cellulose/CuNP com-
posite films reduced by Ocimum sanctum leaf extract. Further,
the peak values are in agreement with those reported by Alek-
seeva et al.[22] in the case of CuNP composites based on cellulose
derivatives. In addition to these four peaks there are several peaks
which may correspond to the diffraction from the constituents of
the leaf extract. The X-ray diffractograms of the composites
 JOURNAL OF MACROMOLECULAR SCIENCE, PART A: PURE AND APPLIED CHEMISTRY 63

Figure 6. (a) Antibacterial activity of matrix and (b) Antibacterial activity of

cellulose/CuNP composites formed using 5mM, 25 mM, 125 mM and 250 mM aq.
CuSO4.5H2O solutions.

Table 1. Inhibition Zone Diameters of matrix and cellulose/CuNP composites. Figure 7. X-Ray diffractograms of (a) matrix and cellulose/CuNPs generated using
different concentrated copper sulphate solutions (b) magnified view of nanocom-
Matrix and cellulose/CuNP composites Inhibition Zone Diameter posite using 250 mM aq. CuSO4.5H2O solution in 2u D 25 to 80 range.

Matrix
5 mMCu
25 mMCu
125 mMCu
250 mMCu
clearly indicate the presence of both CuNPs and Cu2O
nanoparticles.
3.7. TGA
In order to study the effect of CuNPs on the thermal stability of
the composites, the authors carried out the thermogravimetric
analysis of the matrix and the composites. The primary ther-
mograms of the matrix and composites made with different
concentrated aq. CuSO4.5H2O solutions are presented in
Figure 8a. From Figure 8a, it is evident that though the degra-
dation of the matrix was a single step process while in the case
0.7 cm
1.2 cm of the composites it was multi step degradation. For clarity the
1.5 cm
2 cm
derivative thermograms of the matrix and the composite pre-
3 cm pared using 250 mM aqueous CuSO4.5H2O solution were
recorded and are presented in Figure 8b. In the case of matrix
the inflection temperature (the temperature at which degrada-
tion rate is maximum) was found to be 291 C. However, in the
case of the composite, three inflection temperatures at 198 C,
325 C and 462 C were observed. Of these, the degradation
temperature at 198 C may correspond to the possible presence
of trapped/unreacted CuSO4.5H2O.[23] The second degradation
temperature, which is nearer to that of the matrix, belongs to
the decomposition of the matrix component in the composite.
This indicates that the thermal stability of the matrix was
increased by the presence of the CuNPs in the nanocomposite
films prepared. Vyazovkin[23] studied the thermal degradation
of a mixture of copper sulfate and cellulose and observed that
the first degradation temperature of anhydrous copper sulfate

Table 2. Comparative analysis of Anti-bacterial activity of Cellulose/CuNPs composite films using various plant extracts as reducing agents.
S.No Leaf extract used as reducing agent Varying concentrations of metal ion Range of Anti-bacterial activity Zone diameter against E.coli. Ref.

1 Ocimum Sanctum 5 mM, 25 mM, 125 mM and 250 mM CuSO4 8 –18 mm [17]
2 Terminalia Catappa 5 mM, 25 mM, 125 mM and 250 mM CuSO4 2–12 mm [20]
3 Cassia alata 5 mM, 25 mM, 125 mM and 250 mM CuSO4 7 to 30 mm Present Work
64 P. SIVARANJANA ET AL.

Figure 8. (a)Primary thermograms of matrix and cellulose/CuNP composites

formed using different concentrated copper sulphate solutions; (b) Derivative ther-
mograms of matrix and composite made using 250 mMaq.CuSO4.5H2O solution.

took place at a temperature 200 C lower than that of pure cop-

per sulfate. Kett and Price[24] reported the first degradation
temperature of pure anhydrous copper sulfate at 660 C. The
observed degradation temperature of 462 C in the present sys-
tem agrees well with the observations made in both the
cases.[23,24]

3.8. Tensile strength

For packaging and other applications, the composite films have
to possess good tensile strength. In order to examine the effect
of CuNPs on the tensile strength of the cellulose/CuNP com-
posite films, the tensile test for the matrix and the composites
was conducted. The variation of tensile strength with concen-
tration of aq.CuSO4.5H2O solution is presented in Figure 9a.
From Figure 9a, it is evident that the tensile strength of the
composites increased with increasing concentration of aq.
CuSO4.5H2O solutions up to 125 mM and later decreased. This
clearly indicates that CuNPs formed in the composites acted as
reinforcing fillers up to a concentration of 125 mM (the tensile
strength increased from 38MPa of matrix to 70 MPa). The
decrease of the tensile strength for the composites prepared
with 250 mM aq.CuSO4.5H2O solution can be attributed to the
possible agglomeration of the CuNPs at this concentration. The
Figure 9. (a) Tensile strength (b) Tensile modulus and (c) Percentage of elongation
at break of matrix and the cellulose/CuNP composites formed using 5 mM, 25 mM,
125 mM and 250 mM aq.CuSO4.5H2O solutions.
increase in size of the CuNPs (»71 nm for the composites
made using 250 mM aq. CuSO4.5H2O solution) further sup-
ports their agglomeration in some places of the composite. The
tensile modulus and % of elongation at break of the matrix and
composites are presented in Figure 9b and Figure 9c respec-
tively. From Figure 9c, it was observed that in the case of both
the matrix and the cellulose/CuNP composite films, the per-
centage elongation at break was found to exhibit a nonlinear
variation. In order to ensure the difference between the samples
regarding tensile strength, modulus and elongation the statisti-
cal analysis has been performed by one-way analysis of variance
 JOURNAL OF MACROMOLECULAR SCIENCE, PART A: PURE AND APPLIED CHEMISTRY
(ANOVA) in two category: between groups and within groups.
As a part of ANOVA analysis the F-ratio was calculated for all
the cases. To obtain the P-value, F-critical value was deter-
mined from F distribution table for the corresponding degrees
of freedom associated with numerator and denominator values
respectively. In general, if F is equal to or larger than this criti-
cal F-value, then the obtained result is considered as significant
at that level of probability. Accordingly, the result of F-ratio
and F-critical obtained from this work indicates that there is a
statistically significant difference between the mean values
from one type of composite to another at the 95.0% confidence
level.
4. Conclusions
Cellulose pulp was dissolved in environment friendly solvent
(aq.LiOHCUrea) pre-cooled to –12.5 C. The wet cellulose films
in the form of gel were regenerated using alcoholic bath. These
wet films were immersed in 10 wt% Cassia alata leaf extract
and allowed the extract to diffuse into them. In order to in situ
generate copper nano particle (CuNPs), the leaf extract diffused
wet films (matrix) were immersed in different concentrated aq.
CuSO4.5H2O solutions. The change in color of the matrix indi-
cated the in situ generation of CuNPs in the matrix. Apart
from in situ generation of CuNPs in the composites, some
CuNPs were also found to form outside the composite films in
the copper sulphate solutions due to some migration of leaf
extract into the source solutions. The particle size distribution
of the CuNPs formed outside and inside the films was deter-
mined using SEM images. The CuNPs were spherical in shape
with particle size varying from 48 nm to 71 nm in the compos-
ite films. The presence of CuNPs was established by EDX spec-
tra and XRD techniques. No chemical interactions between
CuNPs and the matrix were revealed in the FTIR spectra except
possible electrostatic interactions. The thermal stability of the
matrix component in the composites increased due to the pres-
ence of CuNPs. The cellulose/CuNP composite films showed a
good antibacterial activity against E.coli bacteria. Further, the
composite films possessed good tensile strength. Based on the
antibacterial activity and good tensile strength, the cellulose/
CuNP composite films can be considered for antibacterial
package and medical applications.
Acknowledgments
The authors acknowledge the Institute of Tropical Forestry and Forest
Products (INTROP), Universiti Putra Malaysia, Malaysia for providing
facility for the testing of composite films with the aid of funding through
Putra Grant Vot No. 9441501. Authors also thank the Kalasalingam
University, TamilNadu, India for supporting this research by providing
fabrication facility to the authors.
65
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