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Synthesis and characterization of lignin-poly lactic

acid film as active food packaging material

Ankita Chaubey , Keshaw R. Aadil & Harit Jha

To cite this article: Ankita Chaubey , Keshaw R. Aadil & Harit Jha (2020): Synthesis and
characterization of lignin-poly lactic acid film as active food packaging material, Materials
Technology, DOI: 10.1080/10667857.2020.1782060

To link to this article: https://doi.org/10.1080/10667857.2020.1782060

Published online: 23 Jun 2020.

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MATERIALS TECHNOLOGY
https://doi.org/10.1080/10667857.2020.1782060

Synthesis and characterization of lignin-poly lactic acid film as active food

packaging material
Ankita Chaubeya, Keshaw R. Aadilb and Harit Jhaa
a
Department of Biotechnology, Guru Ghasidas Vishwavidyalaya, Bilaspur, India; bCenter for Basic Sciences, Pt. Ravishankar Shukla
University, Rapur, India

ABSTRACT ARTICLE HISTORY

The present work describes the synthesis, characterisation and antimicrobial activity of lignin- Received 7 February 2020
polylactic acid (PLA) blended film. Film was prepared using co-polymerisation reaction of lignin Accepted 30 May 2020
and lactic acid in the presence of stannous chloride as a catalyst and poly(vinyl alcohol) to get KEYWORDS
the uniform film. The properties of the film was studied by scanning electron microscopy (SEM), Lignin; poly lactic acid (PLA);
X-ray diffraction (XRD), differential scanning calorimetry (DSC) and thermo-gravimetric analysis biodegradability;
(TGA), respectively. The lignin-PLA film demonstrated a significant decrease in swelling ratio antimicrobial activity
and tensile strength in contrast to PLA film. SEM analysis revealed proper blending of lignin
into PLA matrix with slightly rough surface. Film was thermostable and amorphous in nature as
confirmed from the thermal study and XRD analysis, respectively. Further, significant improve­
ment was observed in antimicrobial activity and biodegradability of PLA film after addition of
lignin, which suggest the putative application of lignin-PLA film as the food packaging and
mulch film materials.

Introduction blends, copolymers and to reinforce it with various

inorganic and organic fillers.
Dependence on the petrochemical-based non-
Lignin, the most abundant polyphenolic com­
renewable polymers has become a serious threat to
pound found in biomass, can be used as
the environment. The use of non-degradable poly­
a renewable organic filler. It is the main component
mers from the non-renewable resource is one of the
of the plant cell wall and binds to the cellulose and
major reasons of environmental pollution. Hence,
hemicellulose to provide mechanical support
such problems have driven the efforts to replace the
[1,9-11]. Lignin is widely acceptable for application
petroleum origin polymers with renewable and nat­
in the synthesis of value-added products such as
ure-derived biopolymers [1–3]. Natural resource fil­
resins, films, foams, stabilising agent for nanoparti­
lers are abundant, inexpensive, renewable and fully
cles and nanofibers, it has various functional groups
biodegradable raw materials [4]. Consequently, the
(hydroxyls, methoxyl, carbonyl, and carboxyl) in its
biocomposites with natural resource fillers have
structure [1,3,12,13]. The successful use of lignin in
become a major part of the biodegradable plastic
blends with different biopolymers, such as starch
industry. Poly-lactic acid (PLA) is a linear aliphatic
[14], alginate [1], gelatin [15] and synthetic polymers
thermoplastic polyester which can be obtained from
like poly(vinyl alcohol), poly(ethylene), poly(lactic
lactic acid derived from the fermentation of renew­
acid)and poly(vinyl chloride) have also been reported
able agricultural-based feedstocks [5,6]. The global
in the literature [7,12,16,17]. Moreover, antibacterial
production growth of PLA is envisaged to approxi­
and antioxidant activities of lignin have been
mately quadruple between 2013 and 2020 [7].
reported, which might be helpful to augment the
Furthermore, PLA is biodegradable and have some
applicability of PLA in the food and pharmaceutical
promising properties such as recyclable and compo­
industries [18].
stable with good stiffness and strength, though the
The objective of the current work was to synthesise
applicability of PLA is limited due to high brittleness,
the lignin-PLA blend for its plausible application in
low softening temperature and weak water vapour
food packaging. The synthesised film was charac­
and gas barrier properties, inferior moisture sensitiv­
terised by physicochemically, morphologically and
ity, early physical ageing and poor impact resistance
structurally to evaluate the effect of lignin on the
[2,8, 7]. Therefore, the best way to improve its prop­
PLA matrix. At the end, antimicrobial and biodegrad­
erties and to extend its application field is to prepare
ability properties of the film were investigated.

CONTACT Harit Jha harit74@yahoo.co.in, harit.jha@ggu.ac.in Department of Biotechnology, Guru Ghasidas Vishwavidyalaya, Bilaspur 495009,
India
© 2020 Informa UK Limited, trading as Taylor & Francis Group
2 A. CHAUBEY ET AL.
Material and methods
Commercial aqueous lactic acid (2-Hydroxypropionic
acid), lignosulphonic acid sodium salt, Poly vinyl alco­
hol)-cold water soluble (PVA), Stannous chloride
were purchased from Hi-media Laboratories Pvt.
Ltd, Mumbai, India. Polyethylene glycol-4000 (PEG),
used as a plasticiser was purchased from Qualigens,
India.
Synthesis of poly(lactic acid)-lignin blend
The PLA was synthesised by the method of ring-
opening polymerisation with minor modifications. In
brief, 100 ml lactic acid was heated at 160°C for 2 h.
Further, 1% (w/v) stannous chloride dissolved in
DMSO and lignin (2% w/v) was added to it and kept
at 180°C for 2 h at hot air oven. Stannous chloride
(SnCl2) was used as a catalyst for polymerisation reac­
tion [19]. After 2 h incubation, a viscous blend of
polylactic acid and lignin was obtained.
Preparation of the film
PVA solution (1%) was prepared with 200 ml distilled
water under constant stirring conditions for 2 h under
ambient conditions. Further, the lignin-PLA blend
(0.5%) was slowly added to the PVA solution under
continuous stirring for 20 minutes for homogenisa­
tion. Subsequently, 1% PEG (Poly ethylene glycol) was
added and was kept for another 1 h incubation at 75°
C under continuous stirring condition. Afterwards,
the reaction mixture was removed from the hot plate
and cast onto a plastic tray and left for 3 days at room
temperature to dry. The uniformly dried films were
used for structural and functional characterisa­
tion [20].
Characterisation of the lignin-PLA film
Film thickness
The thickness of the sample was determined using the
manual digital micrometre (Mitutoyo Manufacturing,
Japan) with a precision of 0.001 mm. Ten different
places were measured and the mean thickness was
calculated for each film.
Moisture content
Moisture content (MC) was measured by the method
illustrated by Aadil et al. [21]). Moisture content was
determined by the following equation:
where Mi and Mf are the masses of initial and final
dried samples, respectively. All experiments were car­
ried out in triplicates.
Water solubility and swelling ratio
The water solubility of the film was evaluated accord­
ing to the method described by Nunez-Flores et al.
[22]. Solubility of the film was determined by the
following formula:
where Wi is the initial mass of the expressed film as
dry matter and Wf is the final mass of remained film.
The swelling properties of the films were tested by
the method given by Aadil et al. [23]. Sample (1 × 1 cm
in size) was dipped in 10 ml of distilled water at room
temperature. The weight gain of the swollen film (Ws)
was measured, after blotting the surface with
Whatman No. 1 filter paper. The swelling ratio (SR)
was evaluated using the following equation:
where Ws is the weight of swollen film (g): Wd is the
dry film (g).
Transparency of the film
The light transparency of film was measured at ultra­
violet-visible range (200–800 nm) using a UV-Vis
spectrophotometer (Shimadzu UV-1800, Japan.). The
film was cut into rectangular strips from the various
portions of the film (A, B, C) and pure PLA film was
used as control. Strips of the film were placed in
a spectrophotometer test cell directly and air was
used as a reference. The transparency of the film was
evaluated by the following equation:
where A600 was the absorption at 500 nm, and x is the
thickness (mm) [24].
Mechanical properties analysis
Tensile strength (TS) of the films was determined
according to ASTM standard method D882-01
(ASTM 2001) using a tensile strength and bursting
strength tester (JSR Instruments, Roorkee, India) [21].
Soil degradation
The experiment was carried out in a series of paper
cups, ½ part of the cups was filled with soil. The film
samples were cut into a rectangular shape (1 × 1 cm in
size) and put into cups at 1–2 cm depth from the soil
surface to ensure aerobic conditions for degradation.
The relative humidity was maintained by adding water
periodically. Samples were removed from the soil at
a specific interval of time and weight loss of the film
was calculated by percentage weight loss equation.

where Mi is the initial mass, Mf is the final mass after
an incubation time (t). All the results are the average
of three replicates [25].
Antimicrobial assay
The antimicrobial activity of the prepared pure PLA
and Lignin-PLA films was tested by agar well diffusion
method against the gram-positive bacteria
Staphylococcus aureus (MTCC 96), Bacillus circulans
(MTCC7906), Staphylococcus oralis (MTCC2696) and
Ralstonia eutropha (MTCC2487). The pure culture of
bacteria was sub-cultured on nutrient agar medium.
Bacterial strain was spread uniformly onto the plates
using sterile spreader. A Film solution of 100 mg/ml
was poured into the well of a nutrient agar plate. After
incubation at 37°C for overnight, the diameter of zone
inhibition was measured in centimetre using a scale.
The experiment was performed in triplicates.
Differential scanning calorimetry (DSC) analysis
DSC experiments were performed using a DSC Q20
instrument (Mettler Toledo DSC 822e) under nitrogen
flow at a rate of 30 ml/min. The instrument was
calibrated with a high purity indium standard.
Samples of about 3–5 mg were taken and sealed in
aluminium pans. The sample was heated from 40°C to
250°C at a rate of 20°C/min, then held for 1 minute at
250°C [26].
Thermo-gravimetric analysis (TGA)
Thermo-gravimetric analysis was performed using
a Mettler Toledo (STARe System TGA1 SF/1100)
instrument. The samples were scanned at a heating
rate of 20 K/min from 25°C to 600°C in a nitrogen
atmosphere [27].
XRD analysis
The XRD analysis was performed to study amorphous,
crystalline, or semi-crystalline nature of the polymers
by using an X-Ray Diffractometer (Bruker D2 Phaser)
operating at kV with a Cu-Ka radiation source
(k = 0.154). The diffraction patterns were recorded at
room temperature with the Bragg’s angles (2θ) varying
from 10° to 60° [28].
Table 1. Physical properties of Lignin-PLA blended film.
Thickness Moisture content
Samples (mm) (%)
Control 0.96 ± 0.01 1.10 ± 0.2
LG-PLA 0.16 ± 0.02 1.93 ± 0.49
Values represent mean± standard deviation (SD) of three independent experiments (n = 3).
MATERIALS TECHNOLOGY 3
SEM analysis
The morphology of the PLA and lignin-PLA films was
examined on an SEM instrument (FEI, Nova Nano
SEM 450), at 20 kV accelerating voltage at different
magnifications. The specimens were cryogenically
fractured in liquid nitrogen. The samples were sputter
coated with gold before scanning [17].
Statistical analysis
All assays were performed in triplicates. The analysis
of the films was carried out using Graph Pad Prism
Software Version 5. The results were given as mean
value ±standard deviation (SD).
Results and discussion
Visual appearance
The synthesised lignin-PLA film was uniform, non-
sticky, elastic, and partially soluble in water after 24 h
at ambient temperature. The thickness of lignin-PLA
films was 0.16 mm whereas PLA film was 0.96 mm
thick (Table 1). The control PLA film seemed trans­
parent though the visible appearance of lignin-PLA
film was light brown due to the addition of lignin
(Figure 1).
Moisture content of the film
Moisture content of lignin blended PLA film shown in
Table 1. The percentage of moisture content of lignin-
PLA film was significantly increased up to 1.93%,
which indicates that the addition of lignin into PLA
film increases the moisture content in PLA.
Water solubility and swelling properties
After the 24 h incubation in distilled water, the addi­
tion of lignin into PLA film brought about minor
changes in film solubility. The solubility percentage
of lignin-PLA film was 9.0%, which is slightly lesser to
the control film (9.8%) (Table 1). It was observed that
lignin addition did not affect the solubility of
PLA film.
The swelling ratios of control PLA and lignin-PLA
film are depicted in Table 1. The swelling ratio of pure
PLA films was (323.53%) whereas the swelling ratio of
lignin-PLA was 218.20%, which indicates a significant
decrease in swelling properties of PLA matrix on the
Water solubility Swelling property Tensile strength
(%) (%) (MPa)
9.80 ± 2.40 323.53 ± 41.02 0.562 ± 0.15
9.00 ± 0.81 218.20 ± 55.47 0.495 ± 0.07
4 A. CHAUBEY ET AL.
Figure 1. (a) PLA film (control); (b) Lignin-PLA Film.
addition of lignin. The reduction of swelling ratio in
lignin-PLA film was possibly due to a high degree of
cross-linking between lignin and PLA molecules [23].
Tensile strength
The tensile strength of PLA and LG/PLA film was eval­
uated as it is an important mechanical property of films
and results are shown in Table 1. The tensile strength of
PLA insignificantly decreased on the addition of lignin
which might be due to slightly reduced intermolecular
attraction forces between polymer chains and hence
enhanced the mobility of PLA chains [9,28]. Thunga
et al. suggest that the mechanical properties of the blends
also affected by the molecular weight of lignin 29.
Light barrier property
The results of the transparency of pure PLA and lig­
nin-PLA films are shown in Table 2. In the presence of
lignin, transparency of the lignin-PLA film (0.156) was
significantly increased at wavelength 600nmand
appeared dark in colour which is possibly due to the
chromophoric nature of lignin, which is play impor­
tant role in the protection of UV radiation [30,23].
Soil degradation
During biodegradation visual observations of PLA
blend, film samples were performed. During 10 days
of burial time in soil, the specimens were disintegrated
gradually upon the time course. Some minor charac­
teristic changes such as brittleness and turbidity after
a day of soil burial time were observed in lignin-PLA
film. This might be due to changes in the structure of
Table 2. Light transmittance of lignin – PLA film.
Samples Transparency (A600/X)
Control 0.051
LG_PLA 0.156
the film under fluctuating temperature during biode­
gradation [26]. Moreover, the burial time and humid­
ity some other factors also affected the degradability of
the film. The weight loss (%) of pure PLA film and
lignin-PLA film was determined, as shown in Figure 2.
The films were deteriorated as a function of degrada­
tion time. Lignin-PLA film showed the highest weight
loss initially during the soil burial experiment. Then,
their weight loss decreased slowly until the 8 days of
soil burial time and increased again after 8 days of soil
burial condition. This might be due to the removal of
water molecules from the structure of PLA of the
blended film initially during soil burial. Interestingly,
the soil degradability of lignin-PLA film was greater
than that obtained from pure PLA film.
Antimicrobial assay
After 24 h of incubation, the plates of well-diffusion
experiments were observed for the presence of the
growth inhibition zone around the film loaded wells
and control antibiotics well. The antimicrobial activity
of the lignin-PLA film is shown in Figure 3 (a-b).
Antimicrobial activity was tested against
Staphylococcus aureus (MTCC96), Bacillus circulans
(MTCC7906), Staphylococcus oralis (MTCC2696)
and Ralstonia eutropha (MTCC2487). The film has
shown the zone of inhibition against and gram-
positive B. circulans, which is pathogenic in nature.
Nonetheless, the zone of inhibition was not observed
against S. aureus, gram-negative R. eutropha and
S. oralis. The average diameter of the zone measured
longitudinally and transversely, was around 17 mm
against B. circulans. Previously, the antimicrobial
activity of kraft lignin and lignosulphonate has been
reported [1]. Our study is consistent with the previous
study of Kmita et al.’s study [31], suggesting that the
PLA film shows the antimicrobial activity against
gram-positive bacteria but no antimicrobial activity
against gram-negative bacteria as suggested by [28].
 MATERIALS TECHNOLOGY 5

Figure 2. Weight loss of the film as a function of time of soil burial.

Figure 3. (a) R. eutropha (gram negative) (b) B. circulans (gram positive).

DSC analysis TGA analysis

The DSC heating therograms of pure PLA and
lignin-PLA films are shown in Figure 4. DSC
study suggested that on the addition of lignin the
glass transition temperature of neat PLA
(Tg = 98.29°C) followed by an endothermic event
with ΔH value (475.75 J/g) shifts to higher glass
transition temperature (Tg) at 133.13°C
(ΔH = 271.52 J/g) in the composite (Figure 4a).
In Figure 4(b), some smaller peaks can also be
observed along with the major peak, such peaks­
might be due to the presence of certain micro­
phase separation or immiscibility of hydrophobic
lignin and hydrophilic PLA, which provide free
volume for the PLA molecule mobility [9].
The TGA and DTG scans of pure PLA and lignin-PLA
films are shown in Figure 5(a-b). It is observed in Figure 5
(a) that there are five different steps. The first step ranges
from 59to 100°C, which is possibly due to the moisture
present in the sample. Second peak at Tonset = 174.22°C to
Tendset = 225.64°C which might be due to the degradation
of some low molecular weight molecules. Further steps
show the presence of high molecular weight molecules at
higher temperatures range 400°C to 528°C whereas
Figure 5 (b) shows the initial onset temperature and
final degradation temperature of lignin-PLA from
67.79°C to 476°C. This variation in the thermal degrada­
tion pattern of lignin-PLA indicates the lower thermal
stability of lignin-PLA as compared to pure PLA.
6 A. CHAUBEY ET AL.

Figure 4. DSC analysis (a) PLA film (b) Lignin-PLA film.

Figure 5. Thermo-gravimetric analysis (a) Control film (b) LG-PLA film.


Figure 6. X-ray diffractogram (a) PLA film (b) lignin-PLA film.
XRD analysis
XRD diffraction patterns of pure PLA and lignin-PLA
are shown in Figure 6 (a-b). XRD diffractogram
(Figure 6a) shows a broad diffraction peak for neat
PLA at 2θ = 16.7°which corresponds to pure PLA as
reported in the literature. In Figure 6(b) shows the
shift in diffraction peak of LG/PLA film at 2θ = 15.3°
and around at 19°with a decrease in relative intensity
as compared to pure PLA, which indicates that the
amorphous phase increases in blend films due to addi­
tion of lignin because lignin is itself an amorphous
compound. It is also reported in the literature that due
to the presence of lignin, an amorphous substance, the
crystallinity index of lignin-cellulose nanofibers, and
poly(lactic acid) nanocomposites was limited to only
59% [32].
MATERIALS TECHNOLOGY 7
SEM analysis
SEM studies indicate that the surface morphology
changed severely when lignin added to PLA [33–35].
The development of surface morphology from rough
to smooth with the addition of lignin as shown in
Figure 7 (a-b). The smooth morphology is closely
related to the interaction/coordination between the
two polymers due to cross-linking, which indicates
that the blend is compatible. The fractured surface of
the PLA appears more brittle since less plastic defor­
mation can be detected on the fracture surface of the
pure polymer. When lignin is added to the PLA
matrix, the surface of lignin-PLA showed characteris­
tic changes such as less brittleness and no particle
agglomeration, indicating that a good dispersion of
the filler on the melt was achieved.
8 A. CHAUBEY ET AL.
Figure 7. SEM analysis (a) pure PLA film (b) lignin-PLA film.
Conclusion
In the present research work, lignin-PLA film was
successfully synthesised. The addition of lignin slightly
improves the physical and mechanical properties of
PLA film and shows satisfactory miscibility though
lignin addition also improves the light barrier proper­
ties of PLA film, which could be very valuable for
preventing lipid oxidation in food packaging.
Further, lignin-PLA film also exhibited good antimi­
crobial activity mainly for gram-positive bacteria.
Biodegradability study suggested that lignin-PLA
film was biodegradable and can easily be degraded
using soil in less time period. The study validates the
potential application of lignin-PLA film as packaging
material in the food industry and can also be useful as
mulch film in agriculture.
Acknowledgments
The authors also acknowledge the funding from UGC SAP
project no. F.3-14/2016/DRS-I (SAP-II). The authors are
grateful to Dr Sanjay Lanjhiana, Pharmacy Department,
GGV, Bilaspur, Department of Physics, Pt. Ravishankar
Shukla University, Raipur and Sagar University, Madhya
Pradesh, India for the sample analysis.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
The work was supported by the grants from Chhattisgarh
Council of Science and Technology (CCOST), Raipur, India
(sanction order no: 2142/MRP/2015).
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