International Journal of Biological Macromolecules 52 (2013) 77–84

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International Journal of Biological Macromolecules

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An insight into the properties of Aegle marmelos pectin–chitosan cross-linked

films
Manish Jindal a , Vineet Kumar b , Vikas Rana a , A.K. Tiwary a,∗
a
Department of Pharmaceutical Sciences and Drug Research, Punjabi University, Patiala 147002, Punjab, India
b
Department of Chemistry, Forest Research Institute, Dehradun 248003, Uttarakhand, India

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

Article history: Pectin of Aegle marmelos (AP) ripe fruits processed in equal proportion with chitosan (CH) formed films
Received 6 July 2012 that exhibited minimum swelling index and volume index on exposure to buffers of acidic and alkaline
Received in revised form 11 October 2012 pH. Highest contact angle and spreading coefficient coupled with lowest work of adhesion in all buffers
Accepted 19 October 2012
for this film suggested availability of limited number of functional groups for interaction with water
Available online 26 October 2012
molecules due to optimum cross-linking between NH3 + groups of CH and COO− groups of AP. This
contention was substantiated by the presence of almost negligible charge on this film. The endothermic
Keywords:
transition H characteristic of NH3 + COO− cross-linking between groups in this film was observed
Contact angle
Differential scanning calorimetry (DSC)
to decrease by only 1% after its sequential exposure to pH 1.2 (3 h) and pH 7.4 (6 h). Furthermore, the
Scanning electron microscopy (SEM) absence of pores or erosion in the scanning electron photomicrograph suggested the versatility of this
Spreading coefficient film due to its resistance to acidic and alkaline pH.
Work of adhesion © 2012 Elsevier B.V. All rights reserved.

1. Introduction contribute to the adhesive property of its aqueous dispersions.

Films play a multifacet role in food packaging and pharma-
ceutical products. They are often used for modifying drug release
characteristics from pharmaceutical dosage forms. Films are also
useful in reducing water vapor and gaseous permeation. The prop-
erties of films depend on the type of material used and the process
conditions employed which in turn determine their applications.
Although, synthetic polymers are good film formers and provide
reproducible performance, they lack the benefits of biodegrad-
ability and biocompatibility. Natural polysaccharides, on the other
hand, are well tolerated due to their non-toxic nature and possess
the distinct advantage of being abundantly available, biodegrad-
able, biocompatible and multifunctional due to the presence of
different functional groups. It is due to these properties that natural
polymers offer vast opportunities to the scientist for cross-linking
with other ions/polymers in bid to design new biodegradable mate-
rials for specific applications.
Aegle marmelos fruit is a good source of pectin. The ripe fruits
(commonly called Bael in Hindi language) are edible and are
advocated for use in diarrhea, gastroprotection, ulcerative colitis
and diabetes [1]. The pectins present in A. marmelos fruit (AP)
∗ Corresponding author at: Pharmaceutics Division, Department of Pharmaceuti-
cal Sciences and Drug Research, Punjabi University, Patiala 147002, Punjab, India.
Tel.: +91 9417457385/175 3046304.
E-mail address: aktiwary2@rediffmail.com (A.K. Tiwary).
However, this abundantly available gum is known to possess mod-
erate film forming property that adversely affects its utilization in
food formulation and drug delivery systems. The bael gum con-
tains galactouronic acid indicating the presence of free COO−
moieties. One option for enhancing its application in food and phar-
maceutical industry could be to co-process it with a good film
forming polymer. Chitosan (CH) is a well known, widely inves-
tigated polysaccharidic polymer carrying positive charge due to
presence of NH3 + groups. However, both AP and CH are poor film
formers and readily dissolve in acidic pH when used alone. These
properties of AP and CH limit their applicability in their putative
form.
The presence of COO− in AP and NH3 + groups in CH can be
expected to interact to yield a AP–CH composite complex. Inter-
action between COO− or HSO3 − groups of chondroitin sulfate
[2], sodium citrate [3], sodium alginate [4] with chitosan has been
reported. These films have been investigated for rectification of
mechanical weakness [5,6], decreasing water solubility [7,8] and
improving drug delivery in colo-rectal drug release dosage forms
[9], sustained release drug delivery systems [10] and transdermal
film formulations [11]. On these lines it is logical to contemplate the
interaction of COO− groups present in galactouronic acid moieties
of AP with NH3 + groups present in CH molecules. This interac-
tion can be hypothesized to improve the functional properties of
putative AP and CH as well as film forming property of AP.
Hence, the present investigation was aimed at developing a
interpolymer association between AP and CH and evaluating the

0141-8130/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.ijbiomac.2012.10.020
78 M. Jindal et al. / International Journal of Biological Macromolecules 52 (2013) 77–84
physico-mechanical and electrical attributes of the films formed
by their association.
2. Experimental
2.1. Materials
A. marmelos fruits, fully ripe were collected from local market. AP
extracted from ripe fruits was stored in airtight polypropylene jars
in desiccated condition. De-ionized (Milli-Q) water was used for all
experiments. All other chemicals used were of analytical reagent
grade.
2.2. Extraction of AP
AP was extracted by using a modified method reported ear-
lier [12]. In brief, ripe fruits were collected from the A. marmelos
tree being grown at Punjabi University, Patiala, India campus.
Inner ripe mass was washed and extracted with HCl (1 N) solution
(Scheme 1). Equal quantities of water and extract were added and
boiled for 25 min. The extract was strained through muslin cloth,
cooled, mixed with potassium meta-bisulfite, and filtrate was kept
overnight for clarification. The supernatant was concentrated, pre-
cipitated with alcohol, vacuum dried, and powdered. The pectin
was further purified by dialysis and purified AP was freeze dried.
Scheme 1. Brief outline for extracting pectin from ripe fruits of Aegle marmelos.
2.3. Preparation of CH–AP solution
The total concentration of polymer was fixed at 2% (w/v). CH
was dissolved in 2% (v/v) acetic acid. AP was dissolved separately
in water. Ammonium acetate (5 M) solution was prepared in water
and equally distributed in AP and CH solutions. The CH solution
was then added to AP solution drop wise with continuous stirring
to obtain clear CH–AP solution.
2.4. Preparation of CH–AP film
The CH–AP films were prepared by spraying CH–AP solution
employing spray gun (5 ml/min) with the help of peristaltic pump
using spray gun of 1 mm nozzle (Electrolab, PP20IV, Mumbai, India)
on non-sticky surface of rotating drum with hot air blown at 50 ◦ C
for solvent evaporation. The dried films were microwaved (85 W,
15 s, 5 cycles) to remove bound water from the films. The films were
stored in large size sealed packs till further use.
2.5. Physical characterization of films
2.5.1. Swelling index (SI)
SI [13] of the CH–AP films was determined after exposing the
films to buffers of different pH (1.2, 7.4 or 6.8). The swelling index
was calculated according to the formula:
W2 − W1
SI =
W1
where W1 is the initial weight of the film and W2 is the weight of
the swollen film.
2.5.2. Volume of films (Vf )
Vf in the swollen state was obtained by determining its weight
in air (Wair ) and in n-heptane (Wh ), a non solvent for the CH:AP
films and calculated using the buoyancy principle:
Wair − Wh
Vf =
h
where h is the density (0.684 g/cm2 ) of n-heptane.
2.5.3. Volume index (Vi )
Vi was obtained by determining its volume in air (Vair ) and in
n-heptane (Vh ) and calculated using the buoyancy principle:
Vair − Vh
Vi =
Vh
2.5.4. Film surface contact angle with buffer pH 1.2, 7.4
Contact angle of the prepared films with buffer pH 1.2, 7.4 or
6.8 was determined by employing digital camera (NIKON, 14.2 MP,
10× zoom, in Mega mode). 200 ␮l of respective buffer solution was
placed gently over the film surface with the help of a micro pipette
and the images captured were magnified and printed on A4 paper.
The contact angle of the drop was calculated manually. The results
reported represent mean of twelve determinations.
2.5.5. Work of adhesion (Wa )
Wa was calculated by using the value of contact angle obtained
above:
Wa = L (Cos  + 1)
where  L is the surface tension of the respective buffer calculated
using stalgamometer method.
 M. Jindal et al. / International Journal of Biological Macromolecules 52 (2013) 77–84 79

2.5.6. Spreading coefficient (S)
‘S’ at the buffer film surface was calculated using the equation:
S = L (Cos  − 1)
where  L is the surface tension of the respective buffer calculated
using stalgamometer method.
2.6. Mechanical properties of CH–AP films
The mechanical properties of CH–AP films were studied using
Texture analyzer (TA XT plus, Stable Microsystems, Godalming, Sur-
rey, UK). The observations were recorded in triplicate.
property also favors the use of pectins in edible jellies. However,
moderate film forming ability coupled with its inability to with-
stand dissolution in acidic pH limits the use of putative pectins in
pharmaceutical dosage forms. Similarly, high solubility in acidic pH
limits the use of CH despite its good film forming property. CH con-
tains NH3 + groups in the ionized state whereas, AP is reported to
contain COO− groups. Hence, it would be logical to expect inter-
action between CH and AP due to NH3 + groups of the former
and COO− groups present in the later. This interaction can be
envisaged to yield CH–AP association that would have enhanced
physico-chemical, mechanical and electrical attributes.
3.1. Preparation of CH–AP films

2.7. Thermal characterization of CH–AP films

2.7.1. DSC analysis
CH, AP and CH–AP films prepared by using different CH:AP ratios
were stored in desiccator at 50% RH for 48 h. These films were then
sequentially exposed to buffer pH 1.2, 6.8 or 7.4. All these samples
were then subjected to DSC analysis (40–400 ◦ C) employing heating
rate of 10 ◦ C/min (Lab Sys Evo, Setaram, France).
2.8. Electrical properties
2.8.1. Zeta potential and conductivity
The zeta potential and conductivity of the CH–AP films were
determined by dynamic light scattering (DLS) (Zetasizer Nano ZS,
Malvern Instruments, UK). Each sample was analyzed by placing
the film on a sample cell holder. Three replicate studies were con-
ducted, with three readings for each study. Results are given as the
average ± standard deviation of the experimental values.
2.8.2. Surface morphology
The SEM photomicrographs of freshly prepared CH–AP films and
after exposing them to various pH environments (1.2, 7.4 or 6.8)
were used for evaluating the change in surface morphology (LEO
435VP, Cambridge, UK).
3. Results and discussion
Pectins in general have been known to yield viscous aqueous
dispersions due to their swellability. They are widely used for
this property in confectionary and baked products. Their gelling
The addition of CH solution (in acetic acid) to AP solution (in
water) forms white gelatinous precipitates that do not form films.
CH dissolved in acetic acid bears NH3 + moieties due to ionization.
AP contains galactouronic acid that possesses COO− moieties. The
reaction between NH3 + moieties of CH and COO− moieties of AP
is spontaneous and results in formation of water insoluble gelati-
nous precipitates. Therefore, ammonium acetate was added to both
CH acetate and AP solution for preventing spontaneous interaction
between them. The clear solution obtained after mixing CH and AP
solutions containing ammonium acetate was sprayed over a glass
surface and evaporated at 50 ◦ C. During drying process, ammonium
acetate and excess acetic acid slowly evaporated to form brown
colored, smooth and flexible film. AP alone did not form any film,
whereas, the films formed by using CH alone were brittle and hard.
It was observed that 30:70, 40:60, 50:50, 60:40 and 70:30 ratios of
CH:AP yielded acceptable films, which were subjected to evaluation
of physico-mechanical and electrical properties.
3.2. Physical performance of CH–AP films
Swelling index (SI) is the indicator of amount of water retained
by the film. Vf showed changes in diametric and/or thickness of
films on coming in contact with buffer pH 1.2, 7.4 or 6.8. Vi reflected
proportional changes in film volume parameters with respect to
dried film. Thus, minimum SI, Vf and Vi of APCH5 film formulation
suggested 50:50 proportion of CH and AP to yield films possibly
possessing optimum interpolymer network (Table 1).
The contact angle made by buffer (pH 1.2, 7.4 or 6.8) with a film
surface is indicative of the relative hydrophilic/lipophilic nature of
the film. An increase in contact angle is due to lower wetting and

Table 1
Physical properties of different CH–AP films.

Film code Composition Swelling index (SI) Volume of film (Vf ) Volume index (Vi ) Cos  Work of Spreading
adhesion (Wa ) coefficient

CH AP

Buffer pH 1.2
APCH3 30 70 3.2 ± 0.86 52.61 ± 1.97 13.45 ± 1.05 0.74 ± 0.01 125.15 ± 4.12 −18.44 ± 1.36
APCH4 40 60 2.5 ± 0.52 23.23 ± 1.11 6.95 ± 0.92 0.66 ± 0.05 119.73 ± 4.43 −23.86 ± 1.98
APCH5 50 50 1.5 ± 0.23 14.61 ± 0.81 3.56 ± 0.20 0.20 ± 0.03 86.28 ± 2.19 −57.31 ± 3.79
APCH6 60 40 3.0 ± 0.83 34.97 ± 3.00 9.77 ± 1.07 0.63 ± 0.02 117.12 ± 4.74 −26.47 ± 2.18
APCH7 70 30 3.6 ± 1.05 50.47 ± 3.01 12.64 ± 2.05 0.69 ± 0.02 121.66 ± 4.38 −21.93 ± 2.26
Buffer pH 7.4
APCH3 30 70 4.0 ± 1.09 65.85 ± 5.11 16.39 ± 2.05 0.48 ± 0.01 96.67 ± 3.48 −33.54 ± 2.19
APCH4 40 60 3.5 ± 0.91 73.95 ± 5.02 18.21 ± 2.03 0.40 ± 0.02 91.32 ± 3.96 −38.89 ± 3.17
APCH5 50 50 2.0 ± 0.27 4.95 ± 0.35 1.24 ± 0.25 0.08 ± 0.05 70.77 ± 2.16 −59.44 ± 3.78
APCH6 60 40 3.2 ± 0.84 17.23 ± 1.50 4.53 ± 0.82 0.15 ± 0.02 75.30 ± 3.48 −54.91 ± 3.28
APCH7 70 30 4.2 ± 1.01 41.61 ± 4.01 10.71 ± 1.02 0.41 ± 0.01 92.22 ± 3.85 −37.99 ± 2.96
Buffer pH 6.8
APCH3 30 70 3.7 ± 0.62 25.09 ± 2.00 6.87 ± 1.05 0.40 ± 0.01 80.43 ± 3.60 −34.26 ± 3.75
APCH4 40 60 2.6 ± 0.29 15.87 ± 1.50 4.41 ± 1.11 0.28 ± 0.02 73.84 ± 3.25 −40.85 ± 2.89
APCH5 50 50 1.7 ± 0.26 7.23 ± 0.52 1.83 ± 0.20 0.11 ± 0.02 61.79 ± 2.78 −50.90 ± 3.74
APCH6 60 40 2.9 ± 0.81 31.71 ± 3.02 8.78 ± 1.05 0.14 ± 0.03 65.57 ± 2.94 −49.12 ± 3.58
APCH7 70 30 3.5 ± 1.08 40.16 ± 3.00 10.64 ± 1.05 0.20 ± 0.02 68.64 ± 2.18 −41.05 ± 3.28
80 M. Jindal et al. / International Journal of Biological Macromolecules 52 (2013) 77–84

Table 2
Mechanical and electrical attributes of CH–AP films.

Film code Composition Mechanical Electrical

CH AP Tensile Extensibility (mm) Modulus Zeta potential Mobility Conductivity

strength (N) (N/m2 ) (mV) (cm2 /(V s)) (mS/cm)

APCH3 30 70 4.12 ± 0.23 −22.82 ± 2.12 9155.56 ± 8.65 −41.54 ± 3.17 −3.45e−006 7.15 ± 1.49
APCH4 40 60 4.91 ± 0.32 −28.37 ± 2.43 10,911.12 ± 12.76 −30.83 ± 4.87 −2.40e−004 6.73 ± 0.43
APCH5 50 50 12.12 ± 1.10 −34.19 ± 4.12 26,933.34 ± 16.89 0.27 ± 0.09 2.07e−006 5.29 ± 0.69
APCH6 60 40 10.4 ± 1.32 −26.09 ± 2.18 23,111.12 ± 9.43 3.10 ± 0.42 2.42e−005 7.63 ± 1.30
APCH7 70 30 4.63 ± 0.56 −14.21 ± 1.21 10,288.89 ± 21.28 7.21 ± 1.18 3.14e−006 11.25 ± 1.61

it indicates relatively higher lipophilicity of the film surface [14].
Further, higher work of adhesion (Wa ) indicates stronger bond-
ing between the buffer and film surface. The data summarized
in Table 1 indicates that the film prepared by using equal pro-
portion of CH and AP (APCH5 ) displayed maximum contact angle
as compared to other films, irrespective of the pH of buffer. This
could be attributed to the lipophilic character of the film, which is
in consonance with the lowest SI observed for APCH5 film. These
observations could be attributed to least availability of functional
groups of CH and AP for hydrogen bonding with water because
of optimum interaction between COO− groups of AP and NH3 +
groups of CH during film formation. The contact angle of APCH5 film
was observed to be maximum with buffer pH 7.4, followed by pH
6.8 and least in pH 1.2. The proportion of CH increased while that
of AP decreased from APCH3 to APCH7 films. It is logical to hypoth-
esize that AP would have been present in excess of CH in APCH3
and APCH4 films whereas, CH would have been present in excess
of AP in APCH6 and APCH7 films. AP would be in unionized form in
acidic pH, while CH would be unionized in alkaline pH. The trend
observed for contact angle for all the buffer solutions are justifiable
on the basis of the proportion of CH and AP present in various films
in ionized form.
The work of adhesion determined for different films represents
the affinity of buffers of different pH for these film surfaces. It was
observed that the work of adhesion for APCH5 film was lowest irre-
spective of the pH of buffer. This suggested that the APCH5 film
surface was more difficult to wet as compared to the surface of other
films. Spreading coefficient of all the film surfaces was found to be
negative. These values indicated relatively less hydrophilic charac-
ter of all the film surfaces. In addition, the spreading coefficient for
APCH5 film was observed to be lowest for any buffer. Hence, it could
be suggested that the APCH5 film formed by optimum interaction
of CH and AP was least wettable as compared to other film sur-
faces that probably contained few unreacted functional groups of
CH or AP. The unreacted functional groups of CH and AP could have
been available for interaction with water molecules, thus yielding
relatively hydrophilic surface as compared to the APCH5 film.
3.3. Electrical properties
Zeta potential is an indicator of the net charge present on par-
ticles or films. It is often used in conjunction with mobility and
conductivity to judge the contribution of charged moieties present
in polymeric materials. The data pertaining to zeta potential, mobil-
ity and conductivity of films prepared by employing different
proportions of CH and AP is summarized in Table 2. Film APCH3
containing highest proportion of AP exhibited high negative charge.
The charge on the film became almost negligible when CH and
AP were present in equal proportions (APCH5 film). This could be
attributed to optimum interaction between COO− groups of AP
and NH3 + groups of CH. The charge on films APCH6 and APCH7
steadily increased and was positive possibly due to the presence of
excess CH. Similarly, conductivity was highest for APCH3 film that
contained 30:70 ratio of CH:AP. It was minimum for APCH5 films
containing equal proportions of CH and AP suggesting least avail-
ability of charged functional groups due to optimum interaction
between CH and AP during film formation. It is worthy to note that
there was an inverse relationship between zeta potential and SI. As
the charge on the films increased toward the positive side, the SI
increased. The SI was observed to be minimum when zeta potential
was almost negligible (Fig. 1). The observations suggested that the
presence of ionized functional groups of CH or AP contributed to
high positive or negative charge and resulted in greater SI. Lowest
SI of APCH5 film seems logical due to availability of least number of
ionized groups owing to optimum interaction of COO− and NH3 +
groups.
3.4. Mechanical properties
Adequate mechanical strength and extensibility are basic
requirement of a film for withstanding external stress and main-
taining the integrity as well as barrier properties. Blends of
polysaccharides with different ratios of polymers can be useful
for improving tensile properties. The tensile strength as well as
extensibility of various CH–AP films is summarized in Table 2. Com-
parison of the tensile strength values of films different ratios of CH
and AP (Fig. 2) revealed a three-fold increase in tensile strength of
APCH films when the proportion of CH was increased from 30 parts
to 50 parts. The high tensile strength values of these films could
be attributed to the optimum association between NH3 + groups of
CH and COO− groups of AP. The amino groups (NH2 ) of CH are pro-
tonated to NH3 + in the acetic acid solution, whereas the ordered
structure of AP molecules is destroyed during the solubilization
process. This results in the OH− groups of AP to get exposed and
form hydrogen bonds with NH3 + of CH. However, increasing the
proportion of CH beyond 50% in the films was observed to decrease
the tensile strength of films. Although, no specific reason could
be suggested for this behavior, the enhancement of rigidity due
to increase in the proportion of CH in the films cannot be ruled out.
Extensibility, which is a measure of the maximum distance to which
the film can be stretched before breaking, too showed similar trend.
It increased by 1.5-fold on increasing the proportion of CH from 30
parts to 50 parts and then decreased on further increasing the pro-
portion of CH in the films. It is important to note that the increase in
extensibility was lower in magnitude as compared to the increase
in tensile strength of the films. The modulus of the film summa-
rized in Table 2 supports the correlation that the films containing
maximum cross-linking required more force for breaking it. Film
APCH5 containing 50:50 ratio of AP and CH required greatest force
while the other films required lower force for breaking.
3.5. Thermal properties
The thermogram of pure CH powder indicated one exother-
mic transition at 311.5 ◦ C along with one endothermic transition
at 99.6 ◦ C (Fig. 3A). The exothermic transition is indicative of CH
degradation at 311.5 ◦ C. The occurrence of an endothermic tran-
sition at 99.6 ◦ C could be attributed to the moisture present in
 M. Jindal et al. / International Journal of Biological Macromolecules 52 (2013) 77–84 81

A 4

3.5
Swelling Index (pH 1.2)

2.5

1.5

0.5

0
-50 -40 -30 -20 -10 0 10
Zeta Potential (m V)

B 4.5

3.5
Swelling Index (pH 7.4)

3
2.5 Fig. 2. Mechanical properties of films prepared by employing different proportions
of CH and AP.
2

1.5

0.5

0
-50 -40 -30 -20 -10 0 10
Zeta Potential (m V)

C 4

3.5
Swelling Index (pH 7.4)

2.5

1.5

0.5

0
-50 -40 -30 -20 -10 0 10
Zeta Potential (m V)

Fig. 1. Correlation between zeta potential and swelling index of films prepared by
employing different proportions of CH and AP in buffer pH 1.2 (A), pH 7.4 (B) or pH
6.8 (C).

samples or to the presence of acetic acid moieties. The DSC thermo-

gram of AP (Fig. 3B) showed an endothermic transition at 85.2 ◦ C
and an exothermic transition at 318.6 ◦ C.
The CH–AP (50:50) complex formed by mixing CH dissolved in
acetic acid and AP dissolved in water showed sharp endothermic
transition at 214.1 ◦ C (Fig. 3C). The endothermic transition at 214 ◦ C
observed in this conjugate could have arisen due to carboxylic link- Fig. 3. DSC thermograms of CH (A), AP (B), CH–AP conjugate (C), and films formed
by using equal proportion of CH and AP without exposure (D) and after sequential
age between COO− moieties of AP and NH3 + moieties of CH.
exposure to pH 1.2 for 3 h (E) and pH 7.4 for 6 h (F).
The sharp endothermic transition was evident in thermograms of
all the films prepared by employing different proportions of CH
82 M. Jindal et al. / International Journal of Biological Macromolecules 52 (2013) 77–84

Table 3
Thermotropic attributes of CH:AP films.

Film code Composition Fresh films Exposed to pH 1.2 Sequential exposure to pH

7.4 and 6.8

CH AP Tm (◦ C) H (J/g) Tm (◦ C) H (J/g) Tm (◦ C) H (J/g)

APCH3 30 70 204.14 102.1 194.32 82.1 190.29 57.47

APCH4 40 60 206.98 129 202.76 108.9 201.12 78.12
APCH5 50 50 214.6 184.3 214.12 182.9 214.1 182.4
APCH6 60 40 211.9 117.7 209.3 94.1 201.35 80.41
APCH7 70 30 207.6 97.85 201.1 64.12 190.51 51.76

Fig. 4. DSC thermograms of CH–AP (30:70) films: without exposure (A) and after
sequential exposure to pH 1.2 for 3 h (B) and pH 7.4 for 6 h (C).
and AP (Table 3). It is important to note that lower peak temper-
atures (Tm ) of this characteristic transition at 204 ◦ C and 206 ◦ C,
respectively, were observed for films containing 30:70 or 40:60
ratio of CH:AP. However, the Tm was observed to range from 207 ◦ C
to 214 ◦ C for the films containing 70:30 to 50:50 ratio of CH:AP.
Higher Tm of endothermic transition observed for APCH4 , APCH5
and APCH6 films suggested stronger interaction between CH and
AP in these films (Table 3).
It is essential for a film to withstand exposure to pH 1.2
if it is meant to prevent drug release in the stomach. The
heats of endothermic transitions (H) of freshly prepared films
Fig. 6. DSC thermograms of CH–AP (60:40) films: without exposure (A) and after
sequential exposure to pH 1.2 for 3 h (B) and pH 7.4 for 6 h (C).
containing different ratios of CH:AP were observed to follow the
order: 50:50 > 40:60 > 30:70 > 60:40 > 70:30 (Table 3). The sequen-
tial exposure of APCH5 film containing 50:50 ratio of CH:AP to pH
1.2 (3 h) and pH 7.4 (6 h) resulted in decrease of only 1% in the H
value. Fig. 3D–F does not reveal any marked change in the endother-
mic transition characteristic of cross linking between COO− of AP
and NH3 + of CH after exposure of this film to pH 1.2 and subse-
quently to pH 7.4.
The APCH3 film containing 30:70 ratio of CH:AP (Fig. 4A)
showed 20% and 44% decrease in H of the endothermic transition,

Fig. 5. DSC thermograms of CH–AP (40:60) films: without exposure (A) and after Fig. 7. DSC thermograms of CH–AP (70:30) films: without exposure (A) and after
sequential exposure to pH 1.2 for 3 h (B) and pH 7.4 for 6 h (C). sequential exposure to pH 1.2 for 3 h (B) and pH 7.4 for 6 h (C).
 M. Jindal et al. / International Journal of Biological Macromolecules 52 (2013) 77–84
Fig. 8. SEM images of CH–AP films before and after treatment with various pH environments. ( ) Indicates 100 ␮m.
respectively, after sequential exposure to pH 1.2 (Fig. 4B) and pH
7.4 (Fig. 4C). This film lost its integrity on exposure to pH 7.4.
The loss of H value of APCH4 film (Fig. 5A) containing 40:60
ratio of CH:AP on sequential exposure to pH 1.2 (Fig. 5B) and pH 7.4
(Fig. 5C) was, respectively, 16% and 40%. This film lost its integrity
in pH 7.4. Film APCH6 containing 60:40 ratio of CH:AP (Fig. 6A) lost
20% and 32% of the endothermic H value after sequential exposure
to, respectively, pH 1.2 (Fig. 6B) and pH 7.4 (Fig. 6C). This film too
lost its integrity in pH 7.4. The worst affected film was APCH7 film
containing 70:30 ratio of CH:AP (Fig. 7A). The decrease in H value
of the endothermic transition was 34% and 47% after sequential
exposure to, respectively, pH 1.2 (Fig. 7B) and pH 7.4 (Fig. 7C). This
film could not withstand the exposure to pH 1.2.
These results suggested that shifting of Tm of the endother-
mic transition to higher temperature could not be considered as a
major indicator of the strength of cross-linking. Instead, the mainte-
nance of or negligible decrease in the H value of the characteristic
endothermic transition after exposure of the film seems to be a
83
better indicator of the stability of crosslinked bonds under challeng-
ing environments. Hence, the APCH5 film containing 50:50 ratio of
CH:AP could be suggested to be stable on exposure to pH 1.2 and
pH 7.4.
3.6. Film surface appearance
The SEM images of CH–AP films exposed to pH 1.2 or 7.4 are
depicted in Fig. 8. Smooth texture of freshly prepared films contain-
ing 30:70 or 50:50 ratio of CH:AP was evident. However, the surface
of freshly prepared films containing 40:60, 60:40 or 70:30 ratio of
CH:AP was observed to be irregular. The exposure to pH 1.2 created
holes in films containing 30:70 as well as 70:30 ratios of CH:AP.
Films containing 40:60 or 60:40 ratio of CH:AP were observed to
be relatively unaffected by pH 1.2. However, exposure of all films
except those containing 50:50 ratio of CH:AP to pH 7.4 resulted
in pores as evident in Fig. 8. It is evident that the film containing
50:50 ratio of CH:AP was neither affected by exposure to pH 1.2 nor
84 M. Jindal et al. / International Journal of Biological Macromolecules 52 (2013) 77–84
pH 7.4. It is noteworthy that this film had exhibited least SI among
all the films after exposure to pH 1.2, 7.4 or 6.8. Hence, the film
formed by employing 50:50 ratio of CH and AP could be suggested
to be resistant to the attack of both acidic and alkaline pH.
4. Conclusion
Interpolymer complexation between A. marmelos and chitosan
(50:50) yielded films that exhibited least affinity for acidic and
alkaline buffers. This was manifested in highest contact angle and
lowest SI for this film. The lowest work of adhesion and spreading
coefficient for this film composition too suggested reduced wett-
ability of the film surface. Almost complete interaction between
NH3 + and COO− groups of, respectively, CH and AP was evi-
dent in negligible zeta potential for this film. DSC analysis showed
a sharp endothermic transition at 214 ◦ C for this film composition.
The Tm and H of this endothermic transition remained unaffected
after sequential exposure of the film to pH 1.2 (3 h) and pH 7.4 (6 h).
The surface of this film was not altered after exposure to acidic and
alkaline pH as evidenced from SEM photomicrographs. On conclu-
sion, the results suggest that equal proportions of CH and AP could
be used for preparing interpolymer complexed films that would
be resistant to acidic and alkaline pH and hence, could be further
exploited for use as drug release modulators.
Acknowledgements
The scholarship provided to Mr. Manish Jindal (SRF) by Indian
Council of Medical Research, New Delhi, India, vide sanction order
no. 58/8/2008-BMS to work on this project is acknowledged. The
authors also wish to acknowledge Department of Science and
Technology (FIST grant) New Delhi, India and University Grants
Commission (SAP grant), New Delhi, India for providing the funds
to complete this work.
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