LWT - Food Science and Technology 135 (2021) 109984

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Structure and properties of chitosan films: Effect of the type of solvent acid
Congde Qiao *, Xianguang Ma , Xujie Wang , Libin Liu
State Key Laboratory of Biobased Material and Green Papermaking, School of Materials Science and Engineering, Qilu University of Technology (Shandong Academy of
Sciences), Jinan, 250353, PR China

A R T I C L E I N F O A B S T R A C T

Keywords: The properties of chitosan films can be modified by changing the solvent type, attributing to the different
Hydrogen bonds interaction patterns between chitosan and acids. However, little is known about how these interactions affect the
Ionic interactions structure and properties of chitosan films. In this work, the influence of acid type on the structure and properties
Thermal transition
of chitosan films was studied by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and
Mechanical properties
differential scanning calorimetry (DSC). FTIR spectra showed that the ionic interactions and hydrogen bonding
could occur between chitosan and acid ions. Structural analysis revealed that chitosan was partially crystalline in
hydrochloric and acetic acid films, whereas it was amorphous in lactic and citric acid films. DSC result indicated
that the glass transition temperature of chitosan was much lower in citric acid film with low water content than
in all the other films, suggesting that the citrate ions interacted more strongly with chitosan. A melting transition
appeared in hydrochloric and acetic acid films. In addition, the tensile strength of these films decreased with an
increase in the volume of acid. These observations indicate that the choice of a proper solvent for chitosan may
be desirable for certain special applications.

1. Introduction
Chitosan has been widely used in food, biomedicine and other in­
dustrial fields due to its distinctive advantages of easy processing,
biodegradability, low toxicity, biocompatibility, and availability (Hu
et al., 2020; Liu et al., 2020; Narasagoudr, Hegde, Chougale, Masti, &
Dixit, 2020; Williams, 2011). Nonetheless, the strong inter- and intra­
molecular hydrogen bonds and semi-crystalline structure of chitosan
render it difficult to be dissolved in water and most organic solvents
(Tian et al., 2015). It is generally soluble in dilute acidic solutions below
pH 6.0. The solubility of chitosan is attributed to the protonation of
amino groups on the polymer chains in acid media, resulting in the
destruction of the hydrogen-bonded networks within chitosan molecules
by the electrostatic repulsion between positive charges (Pillai, Paul, &
Sharma, 2009). Parameters such as the degree of deacetylation (DD),
pH, polymer concentration, polymer molecular weight and temperature
are known to influence the dissolution process of chitosan (Rinaudo,
2006).
Generally, chitosan is soluble in acid solutions such as hydrochloric
acid and several organic acids including formic, acetic, butyric, malic,
citric, lactic, oxalic, propionic and succinic acids (Demarger-Andre &
Domard, 1994; Zhong, Song, & Li, 2011). Up to now, acetic acid is the
most frequently used solvent. In chitosan solution, the chain confor­
mation depends on the acidity of the -NH+ 3 (Park, Choi, & Park, 1983).
As a cationic polyelectrolyte, the chain flexibility of chitosan is also
sensitive to the type of counterions (i.e. the acid ions) (Chen, Wang, Hsu,
& Tsai, 2009). Thus the solvent type has an important influence on the
rheological properties of chitosan solutions and chitosan hydrogels
(Lakehal et al., 2019; Soares et al., 2019). In addition, the structure of
chitosan in the solid state is closely related to the structural organization
of the initial polymer solution (Popa-Nita, Alcouffe, Rochas, David, &
Domard, 2010). It has been reported that the chain flexibility is often
manipulated to regulate the properties of chitosan films (Chen, Lin, &
Yang, 1994). Consequently, the solvent acid can alter the structure of
chitosan films, which in turn affect the properties of polymer films. It is
generally observed that the chitosan films prepared from acetic acid
solutions have higher junction density and thus possess higher tensile
strength (TS) and Young’s modulus (E) compared to those cast from
other monocarboxylic acids (Chenni, Djidjelli, Boukerrou, Grohens, &
Saulnier, 2018; Pavoni, Luchese, & Tessaro, 2019; Velásquez-Cock et al.,
2014; Zhong et al., 2011). As compared with films with monoacid sol­
vent, the use of polyacid (malic acid and citric acid) solution as the

* Corresponding author. School of Materials Science and Engineering, Qilu University of Technology (Shandong Academy of Sciences), Postal address: Daxue Rd.
3501, Jinan, 250353, PR China.
E-mail address: cdqiao@qlu.edu.cn (C. Qiao).

https://doi.org/10.1016/j.lwt.2020.109984
Received 28 April 2020; Received in revised form 28 July 2020; Accepted 29 July 2020
Available online 12 August 2020
0023-6438/© 2020 Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
C. Qiao et al.
solvent for chitosan could enhance the mechanical strength of the films,
as a result of potential ionic crosslinking by polyacids (Chen et al., 2007;
Khouri, Penlidis, & Moresoli, 2020). Contrarily, it has been demon­
strated that chitosan films with lactic acid have lower TS and E, due to
the plasticization effect of citrate ions (Park, Marsh, & Rhim; Rhim,
2002). In addition to mechanical properties, other properties such as
water vapor permeability, antimicrobial activity and thermal stability
are also affected by the acid type (Zhong et al.,2011; Pavoni et al.,2019;
Khouri et al., 2020).
The solvent acid is known to interact with chitosan through elec­
trostatic interactions, as well as hydrogen bonds or hydrophobic in­
teractions (Lakehal et al., 2019). All these interactions can influence the
structure and properties of chitosan films. Many studies highlight the
major impact that acids can have on chitosan through electrostatic in­
teractions (Demarger-Andre & Domard, 1994; Khouri et al., 2020;
Velásquez-Cock et al., 2014; Zhong et al., 2011). In fact, this has been
supported by the simple fact that most of the applications of chitosan are
based on its cationic nature. Apparently the size of acids, as counterions,
is expected to affect these ionic interactions between amine and car­
boxylic groups, and thus the properties of chitosan. It has been stated
that the film with larger counterions such as lactate is softer than that
with acetate (Bégin & Van Calsteren, 1999). In the case of interactions
between chitosan and polyacids, hydrogen bonds also contribute
significantly. It has been demonstrated the presence of the hydrogen
bonds between the second oxalic carboxyl group and the non-reactive
parts of chitosan in the chitosan-oxalic acid gel, which play a major
role in its viscoelastic properties (Hamdine, Heuzey, & Begin, 2006).
However, another study indicated that only the ionic interaction pre­
sented in chitosan–oxalic acid membranes (Fadzallah, Majid, Careem, &
Arof, 2014). Additionally, hydrophobic interactions between chitosan
and carboxylic acid could also take place, which could affect the sorp­
tion of carboxylic acids on chitosan (Shamov, Bratskaya, & Avramenko,
2002).
Since the influence of solvents on chitosan is attributed to the
different interaction patterns between chitosan and acids, the informa­
tion about these interactions is critical to understanding the relationship
between the structure and properties of chitosan films. It is reasonable to
assume that the strength of these interactions is closely related to the
structure of acids. Although the effect of the acid type on the chitosan
films has been studied extensively, little is known about how these in­
teractions affect the structure and properties of chitosan films. In this
study, the interactions between chitosan and various acids, including
hydrochloric acid, acetic acid, lactic acid and citric acid, were investi­
gated in details, along with an exploration of structure–property re­
lationships of chitosan films. The aim of this work was to determine the
effect of acid type on the interactions between chitosan and acids, and to
provide a deep understanding of how these interactions impacted this
biopolymer at a molecular level.
2. Experimental
2.1. Materials
Chitosan in the form of powder (90% DD, degree of deacetylation),
hydrochloric acid, acetic acid, lactic acid and citric acid were all pur­
chased from Sinopharm Chemical Reagent Co., Ltd, China. The
viscosity-average molecular weight of chitosan is about 260,000 (Qiao,
Ma, Wang, & | Yao, 2019). All the reagents were used without further
purification. Ultrapure water (18 MΩ cm) was used to prepare various
acid solutions.
2.2. Film preparation and conditioning
Chitosan solutions (2%, w/v) were prepared by dissolving 0.6 g
chitosan powder in 30 mL of 0.15 mol/L hydrochloric, acetic, lactic or
citric acid aqueous solutions. Each film-forming solution was stirred at
2
LWT 135 (2021) 109984
40 ◦ C for 5 h to gain a homogenous solution, and then filtered through a
piece of PTFE membrane with a pore diameter of 0.45 μm to remove
foam and any undissolved impurities. All the film-forming solutions
were casted on a PTFE mold, and dried at ambient temperature for about
48 h, yielding a series of chitosan films.
To perform the conditioning experiment, the as-obtained films were
predried at 50 ◦ C for 24 h in a vacuum drying oven, and then equili­
brated at 25 ◦ C in sealed chambers with relative humidity (RH) of 60%
for at least one week prior to further analysis. In addition, a part of these
conditioned films was heated at 110 ◦ C for 60 min in a vacuum oven.
FTIR and DSC experiments were immediately performed for these heat-
treated samples to evaluate the role of moisture in chitosan-acid sys­
tems. In this work, the films equilibrated at 60% RH and heated at 110
◦
C are noted as hydrated and dry samples, respectively. In addition, the
chitosan films made with hydrochloric, acetic, lactic acid and citric acid
are noted as hydrochloric, acetic, lactic acid and citric acid films,
respectively.
2.3. Fourier-transform infrared (FTIR) spectroscopy
Fourier Transform Infrared (FTIR) data for the films were obtained
by using a Nicolet iS10 spectrometer (Thermo Fisher Scientific Inc.,
Waltham, MA, USA), equipped with an attenuated total reflection (ATR)
accessory. The FTIR spectra were performed from wavenumber
400–4000 cm− 1 with a resolution of 4 cm− 1 and 128 scans. The data
were analyzed with an Omnic software.
2.4. X-ray diffraction (XRD)
XRD patterns of chitosan films prepared from different acids were
recorded by a Bruker.
D8 Advance X-Ray Diffractomer equipped with a multichannel de­
tector by use of a Cu Kα1 (λ = 0.1542 nm) monochromatic X-ray beam.
The relative intensity was recorded in the scattering range of (2θ) 5–50◦
with a scan rate of 1◦ /min.
2.5. Thermogravimetric analysis (TGA)
TGA measurements were carried out on a TGA-1 (Mettler Toledo
Instrument, Zurich, Switzerland) from 30 ◦ C to 800 ◦ C under nitrogen
atmosphere at a heating rate of 10 ◦ C/min.
2.6. Differential scanning calorimetry (DSC)
DSC experiments were performed on a Q 2000 DSC (TA Instrument,
New Castle, USA) equipped with a cooling accessory RCS90. As bio­
logical macromolecules are easy to absorb water to disturb the deter­
mination of thermal transitions, Tzero hermetic pans (withstand an
internal pressure of 3 atm) were used in this work. The hydrated samples
with mass about 5 mg were sealed in the Tzero hermetic pans with lids
and heated in the DSC cell from − 70 to 120 ◦ C at a heating rate of 5 ◦ C/
min. In addition, the dry samples with mass about 5 mg were sealed in
the Tzero pans with lids and heated from − 70 to 180 ◦ C at a heating rate
of 5 ◦ C/min. All measurements were conducted under nitrogen flow of
50 mL/min. The experimental data were determined at least by tripli­
cate on each film sample.
2.7. Mechanical properties
The mechanical properties of chitosan films with different acids were
measured using an Universal testing machine (WDL-005, Jinan Xinshijin
Experimental Instrument Co., China). Test strip dimensions were 50 mm
× 10 mm (length × width). The tensile experiments were conducted at
ambient temperature with a constant deformation rate of 20 mm/min.
The measurements were run three times for each film and took the
average as the results.
C. Qiao et al.
2.8. Statistical analysis
Statistical analyses were carried out using the SPSS software package
computer program (SPSS Statistical Software, Inc., Chicago, IL, USA).
Statistical significance was determined by analysis of variance
(ANOVA). The level of significance was set p < 0.05. All tests were
performed in triplicate.
3. Results and discussion
3.1. Interactions between chitosan and acids
In order to study the interaction between chitosan and acids, FTIR
experiments were performed on chitosan films cast from different acid
solutions. Fig. 1 displays FTIR spectra of the films with different acids.
For all the samples, the broad absorption band between 3600 and 3000
cm− 1 is assigned to the stretching of hydrogen bonded hydroxyl groups
and amino groups of the carbohydrate ring. The peak at 1536 cm− 1 for
the polymer film with acetic acid corresponds to amide II, suggesting the
amino group in chitosan is -NH+ 3 instead of -NH2 (Kumar-Krishnan et al.,
2014). This result confirmed the presence of electrostatic interaction
between chitosan and the acid, and this film is essentially in the acetate
form (Demarger-Andre & Domard, 1994). This peak shifted slightly to a
higher wavenumber of 1552 cm− 1 for film with lactic acid. A similar
observation was reported by Niamsa and Baimark (2009) that the amide
II of chitosan was shifted to higher frequency when the lactic acid was
used instead of acetic acid. The shift of this band indicates the content of
the -NH+ 3 groups is low in lactic acid film. It has been reported that the
interactions between chitosan and the acid increase as the pKa of the
acid decreases (Shamov et al., 2002). As the pKa of lactic acid (~3.86) is
lower than that of acetic acid (~4.8), the ionic interactions between
amine and carboxylic groups are stronger in lactic acid film (Velás­
quez-Cock et al., 2014). Furthermore, the possible hydrogen bonds
formed between the hydroxyl groups of lactic acid and the hydroxyl
groups of chitosan could contribute to the interactions in this system.
The enhancement of interactions between chitosan and acid may result
in a decrease of the concentration of protonated amine in polymer film.
As a consequence, the amide II of chitosan moved to a high frequency.
This phenomenon was more obvious in citric acid film, in which the
amide II became broadened and shifted to a higher wavenumber of
Fig. 1. FTIR spectra of the chitosan films prepared from different acid solu­
tions. All the samples are conditioned at 60% RH. Acetic acid ( ); Lactic acid
( ); Citric acid( ); Hydrochloric acid ( ).
3
LWT 135 (2021) 109984
1560 cm− 1. A plausible explanation for this phenomenon is that the
citric acid possessing two hydroxyl groups and three carboxyl groups
can interact strongly with chitosan through hydrogen bonds and ionic
interactions. Additionally, a strong peak at 1705 cm− 1 appeared, indi­
cating the existence of free carboxyl groups in this film. Similar results
were obtained by Tanigawa et al.(2008) working on chitosan films with
citric acid.
Contrary to carboxylic acids, the introduction of hydrochloric acid
will cause the amide II of chitosan shift to a lower wavenumber of 1519
cm− 1. It can be explained by the fact that hydrochloric acid is a strong
acid and chitosan is fully charged in film whereas it is only partially
charged in all the other films. Moreover, a sharp peak turned up at 1620
cm− 1 in the spectrum of hydrochloric acid film. Obviously, this peak was
not correlated to amide II of chitosan, as the biopolymer had high degree
of deacetylation. It has been demonstrated that the moisture content of
chitosan film with hydrochloric acid is higher than that of films with
carboxylic acids (Demarger-Andre & Domard, 1994; Gartner et al.,
2011). Furthermore, once the sample was heat treated at 110 ◦ C for 60
min, this peak was significantly weakened (Fig. S1). Thus this peak is
very likely to be associated with the residual water that is tightly bound
to the chitosan. A similar observation was also reported by Zawadzki
and Kaczmarek (2010), who attributed this peak to the water physically
adsorbed.
3.2. Crystalline structure of chitosan films with different acids
To investigate the influence of acid type on the structural changes of
chitosan films, the wide X-ray diffraction patterns of the chitosan films
with different acids are compared, as shown in Fig. 2. It is seen that
acetic acid film displays three main diffraction peaks at around 2θ = 12◦ ,
20◦ and 23◦ , respectively, which is associated with the hydrated
conformation of chitosan (Kobaisi, Murugaraj, & Mainwaring, 2012). It
is known that the regenerated chitosan is more amorphous than raw
chitosan due to a partial reconstitution of crystal region of raw chitosan
during the regeneration process (Zhong et al., 2011). The pattern of
hydrochloric acid film was similar to that of acetate sample, except that
the diffraction peaks at around 2θ = 12◦ and 20◦ became stronger for the
former. It has been stated that the XRD pattern of chitosan acetate is very
similar to that of chitosan salts with some inorganic acids, such as HF,
HCl, and H2SO4 (Yamamoto, Kawada, Yui, & Ogawa, 1997). These re­
sults indicate chitosan molecules take up a similar crystalline
Fig. 2. Wide angle X-ray diffraction patterns of the chitosan films prepared
from different acid solutions. All the samples are conditioned at 60% RH. Acetic
acid ( ); Hydrochloric acid ( ); Lactic acid ( ); Citric acid ( ).
C. Qiao et al.
conformation in these systems. In addition, the reflections are more
intense for hydrochloric acid film, suggesting a better organized
long-range structure in this sample than in acetic acid film. However, an
opposite result was reported by Gartner et al. (2011), who investigated
the role of the counterion (acetate vs Cl− ) on the structure of chitosan
films and found that the large acetate counterion with a delocalized
charge plasticized the films enabling a good long range molecular or­
ganization. It should be noted that the concentration of the two dilute
acids used in their work was different ([HAc]~0.17M, [HCl]~0.12M). It
was found that an increase in acetic acid concentration could enhance
the crystallinity of chitosan film (Abdolrahimi, Seifi, & Zadeh, 2018).
Thus an opposite result may be obtained when the two dilute acids are
used at the same concentration.
For lactic acid film, only one broad scattering peak occurred at 2θ =
21◦ , indicating that the molecules in this film were in amorphous state.
Similar results have been reported by other authors (Zhong et al., 2011).
The strong interactions between chitosan and lactic acid may hinder the
movements of the chain segments, and hence restrains the crystalliza­
tion process. Similarly, no crystalline diffraction peak was found in the
diffractogram of citric acid film due to the fact that citric acid could
interact strongly with chitosan. This result was accordance with previ­
ous work by Tanigawa, Miyoshi, and Sakurai (2008).
3.3. Thermogravimetric analysis (TGA)
Fig. 3 displays the TGA curves of chitosan films cast from solutions of
different acids. All the samples show a similar thermal degradation
behavior, which suffer an initial small weight loss at around 120 ◦ C, and
then a sharp weight loss from 190 to 400 ◦ C. The first weight loss at
approximately 120 ◦ C is associated with the removal of absorbed
moisture in the films (Pavoni et al., 2019). For the three films with
carboxylic acids, the weight loss follows the order acetic acid > lactic
acid > citric acid. This result indicates that the moisture content of
samples conforms to the same orders. It is generally accepted that the
water content of the films is related to the degree of protonation of the
chitosan, due to the hydrophilicity of the NH+ 3 groups (Gartner et al.,
2011). From the FTIR results (Fig. 1), it was found that the interactions
between chitosan and acids followed citric acid > lactic acid > acetic
acid. Furthermore, these strong interactions may result in a reduced
charge density of the chitosan chains. This assumption was supported by
Amorim et al. (2016) who demonstrated that citrate anions could be
tightly retained in the electrical double layer of chitosan particles,
partially screening the biopolymer positive charges. Thus, the stronger
Fig. 3. TGA curve of the chitosan films prepared from different acid solutions.
All the samples are conditioned at 60% RH. Acetic acid ( ); Hydrochloric
acid ( ); Lactic acid ( ); Citric acid ( ).
4
LWT 135 (2021) 109984
the interaction between chitosan and acid, the lower water content in
the films. For the film with hydrochloric acid, it failed to accurately
determine the water content from the weight loss at 120 ◦ C, as the loss of
moisture was very limited around this temperature as a result of the tight
binding of water molecules to the chitosan and its Cl− counterion
(Gartner et al., 2011). The notable weight loss from 190 to 400 ◦ C was
related to a complex process involving the decomposition of the acety­
lated as well as deacetylated units of chitosan (Khouri et al., 2020). In
addition, this weight loss may partly correspond to the evaporation of
residual acids in films, which has been suggested from coupled
TGA-mass spectrometry measurements (Quijada-Garrido, Igle­
sias-González, Mazón-Arechederra, & Barrales-Rienda, 2007).
The DTG curves of the samples in Fig. 3 are shown in Fig. S2. It is
seen that there are two main peaks for hydrochloric acid, acetic acid and
lactic acid films. One peak is around 100 ◦ C, corresponding to the water
evaporation, and another peak is related to the decomposition of chi­
tosan, which is around 200, 280 and 280 ◦ C, respectively. For citric acid
film, two overlapping peaks with centers at about 190 and 210 ◦ C may
be due to the evaporation of citric acid derivatives (Khouri et al., 2020),
and the third peak with center at about 350 ◦ C is associated to the
decomposition of chitosan. These observations suggest that the thermal
stability of chitosan films is in the order of citric acid > lactic acid ~
acetic acid > hydrochloric acid.
3.4. Differential scanning calorimetry (DSC)
The glass transition temperature (Tg) is an important parameter of
biopolymers because their physical properties will undergo significant
changes at the glass transition. In this study the influence of acid type on
the Tg of chitosan films was investigated. The DSC thermograms of the
chitosan films with different acids are shown in Fig. 4, and the results are
listed in Table 1. It is seen that the glass transition temperature of acetic
acid film was about 54 ◦ C. This result was almost completely consistent
with Tg values of about 55 ◦ C previously reported for chitosan films
prepared from solutions of acetic acid (Hosseini, Rezaei, Zandi, & Ghavi,
2013; Kumar-Krishnan et al., 2014). It is worthy to note that the glass
transition of chitosan is closely related to the water content of polymer
films. The moisture content of our sample should be comparable to that
of their samples because all films were similarly conditioned at relative
humidities of 50–60%. In addition, both hydrochloric and lactic acid
films exhibit a similar Tg to that of acetic film. In these hydrated
Fig. 4. DSC thermograms of the chitosan films prepared from different acid
solutions. All the samples are conditioned at 60% RH. Acetic acid ( ); Lactic
acid ( ); Citric acid ( ); Hydrochloric acid ( ).
C. Qiao et al. LWT 135 (2021) 109984

Table 1
Effect of solvent system on the glass transition temperature (Tg) and the melting
temperature (Tm) of chitosan films at wet and dry states (1: wet state; 2: dry
state; N: not found).
Solvents Tg1(∘C) Tm1(∘C) Tg2(∘C) Tm2(∘C)
HA 56 ±3 104 ± 3 125 ± 5 N previous work (Velásquez-Cock et al., 2014). In addition, the citric acid
AA 54 ±2 103 ± 2 123 ± 4 N film has the lowest tensile strength and highest elongation at break. As
LA 53 ±2 N 124 ± 4 N the Tg of citric acid film is about 18 ◦ C (Table 1), it exhibits a typical
CA 18 ±1 N 28 ± 2 N rubberlike behavior at room temperature. This rubberlike behavior was
HA: Hydrochloric acid, AA: Acetic acid, LA: Lactic acid, CA: Citric acid.
Values are mean ± SD (n = 3).
chitosan salt films, water molecules are known to act as plasticizers, and
the Tg of chitosan films decreases with an increase in water content. In
addition to moisture, the counterions such as lactate and Cl− also
contribute to the plasticization of chitosan due to their interactions with
polymer (Gartner et al., 2011). The interactions of these counterions
with chitosan were in the order of lactate > acetate > Cl− , whereas the
water content followed a reverse order in these films. Thus the
compromise involving the plasticizing effects of water and counterions
led to a similar Tg for the three samples. For citric acid film, the glass
transition occurred at a low temperature of about 18 ◦ C. A much lower
Tg with values of − 22.5 ◦ C was observed by Tanigawa et al. (2008). It
should be noted that the concentration of citric acid solution used in
their experiments was much higher than that of the acid solution used in
this work. These results suggest that the citrate ions play a major role in
the plasticization of polymer film due to the strong interactions of chi­
tosan with citrate ions.
In the DSC thermogram of acetic acid film, an endothermic peak
appeared at about 103 ◦ C. This thermal event has been demonstrated to
be associated with the melting transition of chitosan (Qiao et al., 2019).
A very similar result was reported by Liu, Zhou, Zhang, Yu, and Cao
(2014) who stated that the Tm of chitosan film with acetic acid was 105
◦
C. It is worthy to note that the determination of the thermal transitions
in hydrated chitosan films is affected by the integrity of the seal of the
DSC pans. The endothermic peak appeared at around 100 ◦ C in her­
metically sealed pans is attributed to the melting transition of chitosan,
whereas it is assigned to moisture evaporation in open systems (Qiao
et al., 2019). Additionally, hydrochloric film displayed a similar Tm to
that of acetic acid film. These observations indicated that chitosan was
partially crystalline in both polymer films, which was confirmed by the
presence of crystalline diffraction peaks in their diffractograms. On the
contrary, the melting behaviors were not observed for lactic and citric
acid films, suggesting that chitosan molecules were in the amorphous
state. This was in accordance with the X-ray spectra shown in Fig. 3,
which displayed completely amorphous patterns.
As the moisture is removed from the polymer films, the Tg will in­
crease accordingly (Fig. S3). Interestingly, hydrochloric, acetic and
lactic acid films also display a similar Tg with values of about 123 ◦ C,
much higher than that of the hydrated samples. However, there is a little
increase of Tg for citric acid film, which varied from 18 to 28 ◦ C. These
results indicate that only citrate ions act as plasticizers in the dry chi­
tosan salt films. In addition, the melting transition was not observed in
these films. In the absence of moisture, chitosan usually undergoes
decomposition prior to melting due to its rigid-rod polymer backbone
with strong inter/-intrachain hydrogen bonds (Murray & Dutcher,
2006).
highest tensile strength and lowest elongation at the break, a typical
pattern of brittle materials. Similar results were observed for the chi­
tosan film made with acetic acid (Suyatma, Tighzert, Copinet, & Coma,
2005). Compared to acetic acid film, the lactic acid film has a low tensile
strength and high elongation at the break. This result was in accord with
frequently observed in chitosan-citric acid systems (Park, Marsh, &
Rhim, 2002; Bégin, &; Tanigawa et al., 2008; Bégin and Van Calsteren,
1999).
In the present work, the tensile strength of these films was found to
be in the order of acetic acid > lactic acid > citric acid, which decreased
with the molecular weight of the acids. A similar correlation was found
between Young’s modulus and volume of acid for chitosan films (Bégin
& Van Calsteren, 1999). It could be explained in term of the interactions
between chitosan and acids. Compared to acetate and lactate ions, the
citrate ions could interact more strongly with chitosan, and the inter­
chain and intrachain hydrogen bonds in chitosan were destroyed,
resulting in a lower mechanical strength. In the case of lactic acid and
acetic films, although their Tg are similar, their aggregation structures
are different. The former is amorphous, whereas the latter is partially
crystalline and thus has a higher mechanical strength. These observa­
tions indicate that the mechanical properties of the chitosan films are
quite different depending on the counter ions.
4. Conclusions
Interactions between chitosan and acid ions depend on the nature of
acids. In hydrochloric and acetic acid films, the ionic interactions occur
and the chitosan is partially crystalline. While in lactic and citric acid
films, the ionic interactions as well as hydrogen bonding exist and the
biopolymer is amorphous. The hydrochloric, acetic and lactic films
exhibit similar Tg values at around 54 ◦ C, whereas a lower glass tran­
sition occurs at 18 ◦ C in citric acid film. Furthermore, a melting transi­
tion of chitosan is observed at about 103 ◦ C in hydrochloric and acetic
acid film. However, this thermal transition is absent when the moisture
is removed from the sample films. The stress-strain curve of acetic acid
film shows a typical pattern of brittle materials, while the citric acid film
shows characteristic features of rubber-like materials. In addition, the

3.5. Mechanical properties

The influence of acid type on the mechanical properties of chitosan

films was investigated. The stress-strain diagram of hydrochloric acid
film was not shown since it could not be tested due to its extreme brit­ Fig. 5. The stress-strain curves of the chitosan films prepared from different
tleness. Fig. 5 exhibits the stress-strain curves of the chitosan films with acid solutions. All the samples are conditioned at 60% RH. Acetic acid ( );
different acids at room temperature. The acetic acid film presents Lactic acid ( ); Citric acid ( ).

5
C. Qiao et al.
lactic acid film exhibits an intermediate behavior. These results suggest
that it may be possible to improve the performances of chitosan films by
the choice of a proper solvent.
CRediT authorship contribution statement
Congde Qiao: Conceptualization, Supervision, Data curation,
Formal analysis, Validation, Visualization, Writing - original draft,
Writing - review & editing. Xianguang Ma: Methodology, Investigation,
Formal analysis, Writing - review & editing. Xujie Wang: Investigation,
Writing - review & editing. Libin Liu: Formal analysis, Writing - review
& editing.
Declaration of competing interests
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgement
The work was supported by the Foundation (No. ZZ20190209) of
State Key Laboratory of Biobased Material and Green Papermaking, Qilu
University of Technology, Shandong Academy of Sciences.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.lwt.2020.109984.
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