Polymer xxx (xxxx) xxx

Contents lists available at ScienceDirect

Polymer
journal homepage: http://www.elsevier.com/locate/polymer

Structural, thermal, and mechanical properties of gelatin-based films

integrated with tara gum
Luca Nuvoli a, Paola Conte b, Costantino Fadda b, José Antonio Reglero Ruiz c, José M. García c,
Salvatore Baldino d, Alberto Mannu d, *
a
Department of Chemistry and Pharmacy, University of Sassari and INSTM, Via Vienna 2, 07100, Sassari, Italy
b
Department of Agriculture, University of Sassari, Viale Italia 39/A, 07100, Sassari, Italy
c
Departamento de Química, Facultad de Ciencias, Universidad de Burgos, Plaza de Misael Bañuelos s/n, 09001, Burgos, Spain
d
Dipartimento di Chimica, Università di Torino, Via Pietro Giuria, 7, I-10125, Torino, Italy

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

Keywords: Films with different morphology can be obtained by mixing fish gelatin, Tara gum and glycerol in different ratio
Tara gum and by subjecting the Tara gum to a ball milling treatment before its use. The amount of the plasticizer glycerol,
Gelatin fish as well as the type of Tara gum employed (as received or milled) resulted, from SEM and AFM analyses, to strong
Biofilms
influencing the morphology of the films and their density. Also, the morphological differences determine
different thermal and mechanical behaviours. In particular, the employment of milled Tara gum allows to
improve the thermal stability, as well as the mechanical properties of the polymers. A similar outcome can be
obtained by increasing the glycerol content, which can be used up to 20 wt%. Glycerol amounts exceeding that
percentage, are detrimental for the quality of the films and reduce their thermal and mechanical performances.

1. Introduction such as drug delivery or tissue engineering [12,13]. Of particular in­

terest, especially for the packaging and biomedical sectors, are the Fish
During the last years, fervent research activities have been conducted Gelatin (FG) based films. FG is cheap, biodegradable and easy to process
on the engineering of composite gels. The possibility to tune gel prop­ for making films. In addition, its mechanical and optical properties can
erties combining specific components has gained increasing importance be modulated by adding polyols which act as plasticizers. The main role
in many application fields, including food, drug delivery, tissue engi­ of such additives consists in the denaturation of the FG’s proteins by
neering and wound dressing [1–3]. reducing the interactions between protein chains and modifying the
Among the several components usually employed for composite gels secondary, tertiary and quaternary structures [14]. Within the polyols
engineering, gelatin, a mix of denaturated proteins derived by collagen commonly employed for such purpose, glycerol stands out as the ideal
[4–6], have attracted particular interest for biomedical and food pack­ candidate. It showed good ability to interact with FG, eco-compatibility,
aging sectors, due its remarkable properties. As a matter of fact, gelatin and it is available in large amount as by-product of waste vegetable oils
is biocompatible, biodegradable, shows high capacity of cell adhesion treatment [15], e.g. from biodiesel production [16]. On the other hand,
and migration [7], shows antimicrobial properties [8], and it has been the use of fish gelatin in composite films is still limited by several factors
successfully exploited for tissue engineering [9,10]. Thermal properties such as low stability, poor mechanical strength and low elasticity [17].
of gelatin have been characterized in different papers. For example, Many studies have been then conducted with the aim to solve these
Rahman et al. [11] presented a complete study about the glass transition weaknesses by exploring the doping with different additives including
and melting points of commercial mammalian gelatin, tuna gelatin, synthetic and biological polymers [18].
bovine gelatin and porcine gelatin. Glass transitions temperatures varied Although different studies have been performed on the use of FG in
from 23 to 75 ◦ C, whereas melting points were measured between 115 composites in combination with various additives, the interaction be­
and 190 ◦ C. These wide range of values, combined to their biocompat­ tween gelatin and different natural gums in modifying the thermal and
ibility, make them suitable in many different biomedical applications, mechanical properties of gelatin has been only recently explored [19].

* Corresponding author.
E-mail address: [email protected] (A. Mannu).

https://doi.org/10.1016/j.polymer.2020.123244
Received 16 July 2020; Received in revised form 13 October 2020; Accepted 16 November 2020
Available online 24 November 2020
0032-3861/© 2020 Elsevier Ltd. All rights reserved.

Please cite this article as: Luca Nuvoli, Polymer, https://doi.org/10.1016/j.polymer.2020.123244

L. Nuvoli et al. Polymer xxx (xxxx) xxx

Natural gums have shown to allow structural engineering of thin films Table 1
while guaranteeing the biodegradability and edibility of the material Nomenclature, density and composition of the films fabricated.
[20]. Specific combinations between natural gelatin, glycerol and nat­ Film FG (eq. Ta (eq. Ball Gly (wt Density (g/
ural gums have found important application especially in food pack­ wt.) wt.) Milling %) cm3)
aging [13]. TGG01 1 1 No 20 7.9
Regarding the most employed natural gums (Tara, guar, and locust TGG02 1 1 Yes 20 8.6
beam), Tara gum (Ta) shows intermediate water solubility with respect TGG03 1 1 No 40 6.8
to guar (cold soluble) and locust beam gum (cold insoluble), making it TGG04 1 1 Yes 40 8.2
TGG05 1 1 No 60 6.5
ideal e.g. as food hydrocolloid [15]. Tara gum is obtained by grinding TGG06 1 1 Yes 60 7.2
the endosperm of the seeds of Caesalpinia spinosa (Fam. Leguminosae) TGG07 2 1 No 20 7.8
and it is composed by polysaccharide chains of high molecular weight TGG08 2 1 Yes 20 8.7
based on the galactomannan unit (See scheme S1 of the Electronic TGG09 1 2 No 20 9.1
TGG10 1 2 Yes 20 9.8
Supplementary Information file, ESI). It has been approved as food ad­
FG/ 1 0 No 20 4.7
ditive due to its non-toxicity and thus it has gained value as film additive Gly
[21]. When employed in gel composites, tara gum acts as thickening Ta/Gly 0 1 No 20 10.0
agent and stabilizer.
In this context, herein, the fabrication and characterization of novel
films based on fish gelatin (FG), Tara gum (Ta), and glycerol (Gly), is different techniques. First, SEM images were taken using a FEI Quanta
reported. Ten films containing different ratio of FG/Ta, different glyc­ 200 scanning electron microscope. The samples were placed on a
erol content and different types of Ta, are described and characterized. double-sided carbon tape and examined at an acceleration voltage of 20
For each film the structural, thermal, and mechanical properties are kV under high vacuum. Also, Atomic Force Microscopy (AFM) images
discussed by meaning of, respectively, IR, X-ray, and SEM, Thermog­ were taken at RT using a confocal AFM-RAMAN model Alpha300R –
ravimetry (TG) and differential scanning Calorimetry (DSC), and tensile Alpha300A from WITec, using an AFM tip of 42 N/m.
properties analyses. The thermogravimetric analysis data were recorded on a TA Instru­
ment Q50 TGA analyzer. TGA tests were performed under O2 (synthetic
2. Materials air) atmosphere using the next procedure: first, samples were heated
from RT to 100 ◦ C at 10 ◦ C/min, and then kept during 10 min to elim­
Tara gum (Aglumix 01, particle size 149 μm) and Fish Gelatin inate the moisture content. Then, samples were heated up to 800 ◦ C at
(LapiFish, particle size 2380 μm) were purchased, respectively, from 10 ◦ C/min.
Silvateam Food Ingredients S.r.l. and Lapi gelatin S.p.a. Fish gelatin with DSC analyses were performed using a DSC Q200 TA Instruments
the following characteristics was employed: Bloom 280, mesh size 8–70, equipment. Samples were tested using a four-cycle procedure [25]. In
protein amount between 85% and 90%, water content 10–12%, and salts the first cycle, after 5 min of stabilization at RT, samples were heated up
content 1–2% [22]. to 150 ◦ C at 10 ◦ C/min and then stabilized during 5 min before cooling
Glycerol (90%) was purchased from Analyticals Carlo Erba, while down in the second cycle to 30 ◦ C at 20 ◦ C/min. In the third cycle,
acetone (>99%) and ethanol (>99.5%) from VWR. All chemicals were samples were heated up to 150 ◦ C at 10 ◦ C/min and after 5 min of
used as received without any further purification. stabilization at 150 ◦ C, the fourth cycle was performed: samples were
cooled down to 30 ◦ C at 20 ◦ C/min. All the tests were performed under
3. Experimental equipment and methods N2 atmosphere (flow rate 50 ml/min). Mass of the samples was fixed at
approximately 20 mg in each test. Glass transition temperature (Tg) was
Fourier transform infrared (FTIR) analysis was performed using a determined in the third cycle.
Bruker infrared Vertex 70 interferometer. The spectra were recorded on To determine the tensile properties of the films, strips of 5 mm in
the films in transmission mode, in the 400-4000 cm− 1 range by aver­ width and 35 mm in length were cut from each film. Tensile tests were
aging 64 scans with 4 cm− 1 of resolution. carried out on a SHIMADZU EZ Test Compact Table-Top Universal
Principal Component Analysis (PCA) as well as Partial Least Squares Tester at 20 ◦ C. Mechanical clamps were used and an extension rate of 5
Discriminant Analysis (PLS-DA) were conducted using of the tool mm/min was applied using a gauge length of 9.44 mm. At least 4 strips
Metaboanalyst 4 [23]. The spectral data were centered and auto-scaled were tested for each film in order to calculate the average value for each
before the PCA and PLS-DA analysis. parameter determined.
The XRD patterns were collected using a Rigaku SmartLab X-ray
powder diffractometer aligned according to a Bragg–Brentano geometry 4. Preparation of films
with Cu Kα radiation (λ = 1.54178 Å) and equipped with a graphite
monochromator in the diffracted beam. Since the polymers showed The preparation of gelatin-based films is here detailed and schema­
broad haloes typical of amorphous condition, it was deliberately tized in Fig. S1 of the ESI. Tara powder was added to a solution of
assumed to restrict the reciprocal space investigation in the angular glycerol and distilled water and the mixture was stirred at 85 ◦ C during
range from 5◦ to 80◦ in 2θ, which allows to determine the main shape 30 min until a homogeneity was reached (I). Then, Fish gelatin was
features of specimen. Powders have been deposited in an amorphous added at 85 ◦ C and stirred until dissolution (II). The mixture was sub­
glass sample holder for measurements. jected to ultrasound for 15 min to remove air bubbles (III). Film-forming
Ball milling of the Ta powders (15 g) was carried out in a SPEX solutions were poured into Steriplan® petri dished (80 mm × 15 mm)
Mixer/Mill 8000 at a rotation speed of 875 rpm during 1 h, using two and dried at room temperature for 6 h and then located at 50 ◦ C in
zirconia balls of 2 g each one. Taking in consideration the very low humidity-controlled oven for 16 h (IV). The corresponding film was
amount of powders which are usually trapped (1 mg) at each impact, thoroughly washed with ethanol and acetone to afford the desired gel.
and the stochastic nature of the ball milling process, over 200k colli­
sions, occurring under the mechanochemical conditions above reported, 5. Results and discussion
should be enough for refining homogenously all the batch powders [24].
Then, two different Tara gum powders were obtained: as received (or 5.1. Density and visual aspect of the films
non-milled) and milled.
The morphologies of the powders and films were observed by Ten films composed by Fish gelatin (FG), Tara powder (Ta) and

2
L. Nuvoli et al. Polymer xxx (xxxx) xxx

data was conducted. The normalized frequencies relative to films

TGG01-TGG10 were considered for building a spectral fingerprint of
each film. The aim of such statistical analysis was to distinguish between
films containing milled and not-milled Tara. At first, non-supervised
Principal Component Analysis (PCA) was conducted, as shown in
Fig. S3 of the ESI.
As expected, PCA analysis of the FT-IR intensities doesn’t allow to
reach a perfect discrimination. In fact, unbiased PCA method works
properly when within-group variation [30], in this case related with
FG/Ta ratio and glycerol content, is sufficiently less than between-group
variation (milled and not-milled Tara). Nevertheless, a supervised
approach as the Partial Least Squares Discriminant Analysis (PLS-DA)
Fig. 1. Photographs of the FG/Gly, Ta/Gly, TGG01 and TGG02 films. can be used for classify the two considered groups and discuss properly
differences between films, as it is presented in Fig. S4 of the ESI, in which
glycerol (Gly) were prepared with different Ta/FG ratio (1/2, 1/1, and 2D Component 1 vs Component 2 Plot of FT-IR intensities of films
2/1) and Gly amount (20, 40, and 60 wt%). Additionally, two reference TGG01-TGG10, containing milled (M) and not-milled (NM) Ta are
films (FG/Gly and Ta/Gly) were also prepared to compare their prop­ shown.
erties with the FG/Ta/Gly films. Finally, two types of Ta were consid­ From the information of Fig. S4 it is possible to appreciate the
ered: as received by the purchaser, and milled. Nomenclature, density discrimination between films contained milled and not-milled Ta ob­
and composition of the films fabricated are reported in Table 1. tained through multivariate PLS-DA analysis. The analysis, even quali­
Table 1 shows that densities obtained vary from 6.5 g/cm3 for tative, of the distances between the labels in the Scores Plot is of
TGG05 film to 9.8 g/cm3 for TGG10 film. Also, it can be noticed that the particular interest. Comparing films containing milled (M) and not-
density of the as-casted films increases for systems containing milled milled (NM) Tara, it is possible to notice incremental differences
Tara powder compared to those produced using the non-milled one following the trend TGG03-TGG04, TGG05-TGG06, TGG07-TGG08, and
(TGG02 vs TGG01). A relevant influence glycerol content on the density TGG09-TGG10, indicating an increasing effect of the Ta grain size which
values was observed, which decrease at higher wt% of glycerol (TGG06 depend on the amount of glycerol and on the FG/Ta ratio. More glycerol
vs TGG02). As expected, pure FG film shows the lower density value is present, more the milled Tara is distinguishable from the not-milled
(4.7 g/cm3), while pure Tara film presents the higher density value one (TGG02, TGG04, TGG06 vs TGG03, TGG05, and TGG07). As mat­
(10.0 g/cm3). ter of fact, films containing an FG/Ta ratio of 1/2 or 2/1 seems to be
From a qualitative visual analysis, film specimens were transparent more sensitive to the type of Tara (TGG08, TGG10 vs TGG09, and
despite Ta/Gly presented a slightly yellow to brown appearance with TGG10). FT-IR qualitative analysis highlighted important chemical dif­
respect to FG/Gly film. On the other hand, in TGG01 to TGG10 films ferences within films with different glycerol content and Tara typology.
opaqueness is reduced. This means that opacity could be related to the If the effect of the plasticizer (glycerol) it is known and expected, the
Tara adding effect and its dispersion during film preparation, causing possibility to affect the composition of a film by changing the gran­
the reduction of the film transparency with respect to starting Fish ulometry of the Tara deserves more attention.
gelatin film. This characteristic could limit the use of films composed by
only Tara in food packaging applications. In Fig. 1 the photographs of 5.3. X-ray diffraction analysis
the films FG/Gly, Ta/Gly, TGG01 and TGG02 are shown.
With the aim to better assess the structural differences between films
5.2. FT-IR analysis TGG01-TGG10, X-ray diffraction analysis was conducted. Fig. S5 of the
ESI displays the XRD patterns of all the films prepared. Ta presents a
A first quality check on the effect of the preparation procedure on the broad peak at ~19◦ indicating amorphous and crystalline regions exis­
chemical composition of the constituents of films TGG01-TGG10 was ted because a large amount of –OH groups interacted via intermolecular
performed by FTIR analysis. Results are displayed in Fig. S2 of the ESI. hydrogen bonds. This peak partially disappeared in the composite film,
The absorption bands for gelatin-based composite films in the IR spectra indicating that a part of Tara molecules was in ordered arrangement,
are situated in the amide band region. The band situated around 3299, which was interrupted in the grafting process; therefore, the obtained
1635, 1550, 1238 cm− 1 correspond to amide-A (NH-stretching coupled composite is amorphous. After grafting, the peak strength decreased,
with hydrogen bonding) and water molecules, amide I (C– – O stretching/ which indicated that intermolecular hydrogen bonds were damaged.
hydrogen bonding coupled with COO), amide II (bending vibration of The diffractogram pattern acquired on the FG/Gly film was typical of
N–H groups and stretching vibrations of C–N groups), and amide III a partially crystalline gelatin with a sharp peak located at 2θ = 7.1◦
(vibrations in the plane of C–N and N–H groups of bound amid). The (d101 = 12.29 Å) and a broad peak located at 2θ = 20◦ (d101 = 4.0 9 Å),
bands in the FTIR spectrum of Tara powder appear in two major regions, as shown in Fig. S5. These characteristic peaks are usually assigned to
3700-2500 cm− 1 and 1700-700 cm− 1. The broad band at 3700-3000 the triple-helical crystalline structure in gelatin. In particular, the first
cm− 1 is a result of O–H stretching vibration which is associated with diffraction peak at 7◦ (sharp and intense) is directly related to the
free, inter and intra-molecular bonded hydroxyl groups. The shoulder- diameter of the triple helix. It was also found that the addition of polyols
shaped band at about 2920 cm− 1 is the stretching vibration of –CH2. such as glycerol most often decreases the intensity of the first peak (2θ =
The bands at 1020 cm− 1 is characteristic of the O–C stretching vibration 7◦ ). Furthermore, it has been observed that the addition of tara gum
of the anhydro glucose ring. These bands indicate that Tara powder has shifted the diffraction angle from 7.1◦ to 9.12◦ (for TGG05/06 and
the general properties of a polysaccharide [16,26]. TGG09/10 films) then decreasing the diameter of the inter reticular
If from one side the FT-IR data confirm the expected composition of triple helix. On the other hand, this intensity of peaks decreases and, in
the films, ruling out any thermal decomposition which can occur during some cases, completely disappeared (TGG01/02, TGG03/04 and
the synthesis from the other side [16,27], by a visual analysis of the plots TGG07/08 films). The disappearance of X-ray diffraction peaks corre­
reported in Fig. S2, few information can be obtained on the differences sponding to the composites films confirms the interaction between two
between films with different composition [28,29]. biopolymers. This phenomenon illustrates the reduction hydrogen
In order to analyse better the available FT-IR data and to extrapolate bonds between hydroxyls group of gelatin and those of anhydroglucose
as more information as possible, a multivariate analysis of the spectral of Tara gum, which limited the movement of molecules and thus

3
L. Nuvoli et al. Polymer xxx (xxxx) xxx

Fig. 2. SEM images of the films prepared (bar scale 100 μm).

prevented crystallization. homogeneous although pores were observed on the top of each system,
The addition of glycerol seems to decrease the intensities of the except in the case of FG/Gly film. In addition, bigger and fewer pores
gelatin peak making the film more amorphous as compared to control were found in the TGG01, TG003, TGG05, TGGO7 and TG009 films, all
gelatin film. This is probably due to the high stability of these films when of them fabricated using the un-milled Ta powder. On the other hand,
glycerol was added. Finally, a clear effect of BM was observed only on more and smaller uniform pores were observed on the surface of the
TGG06 system which presented a significant decrease on the peak in­ TGG02, TG004, TGG06, TG008 and TGG10 films, in which ball milled
tensity at 2θ = 19.9◦ with respect to TGG05. This effect could be related Tara powders were introduced. Then, it is clear that this difference could
with the synergic effect of glycerol addition and Tara gum added with be ascribable to the reduced Tara particles sizes obtained upon ball
refined particle size, and it confirms the influence of Tara granulometry milling.
qualitatively observed by FT-IR. Fig. 4 presents the AFM surface images of TGG09 and TGG10 films,
in which the distribution of Tara powder (non-milled in the case of
5.4. Morphological analysis TGG09 film and milled in the case of TGG10) is observed. AFM surface
images do not show great differences in the Tara powder distribution,
An exhaustive morphology analysis of the films TGG01-TGG10 was showing that, in general, Tara powder is well dispersed in the film
conducted by different approaches. Firstly, Scanning Electron Micro­ surface. Only a few agglomerated Tara particles are observed in the
scopy (SEM) images of the films were collected as shown in Fig. 2. surface of the films, as it can be seen in the yellow circles spotted of
Micrographs in Fig. 2 show that surfaces of TGG films were Fig. 4a and c.

4
L. Nuvoli et al. Polymer xxx (xxxx) xxx

Fig. 4. AFM surface images of TGG09 (a and b) and TGG10 (c and d) films.

The effect of the milling ball can be analyzed from two different
points of view. First, it seems that the size of the agglomerated particles
is slightly higher when non milled Tara is employed, as it can be seen in
the agglomeration of Tara particles in Fig. 4a (TGG09, non-milled Tara)
and 4c (TGG10, milled Tara). Secondly, the size distribution of Tara
particles can be observed in the AFM surfaces images presented in
Fig. 4b (TGG09) and 4d (TGG10). In this case, there is not a very clear
difference between the particles size of non-milled (Fig. 4b) and milled
Tara (Fig. 4d), but it can be detected that milling the Tara powder could
reduce the particle size compared to non-milled Tara. In this sense,
Fig. 4d show Tara particles with sizes below 1 μm of diameter, and on
the other hand, Fig. 4b presents, in the bottom-left zone, Tara particles
with higher sizes (around 1 μm).

5.5. Thermal stability of films

Preliminary structural and morphological analyses on films TGG01-

TGG10 revealed a not negligible effect of Tara granulometry. Films with
different structures should manifest different thermal and mechanical Fig. 5. DSC curve (third cycle) obtained for TGG02 film.
behaviour. Regarding the thermal stability, it was assessed by TGA
analysis. Information about the degradation and mode of decomposition Additionally, Differential Scanning Calorimetry (DSC) analysis were
under the effect of heat were also acquired. In Fig. 4, the TGA thermo­ performed on TGG01-TGG10 films. The DSC measurements were
gram of film TGG02 is presented as an example. employed for the determination of the glass transition temperature (Tg),
Fig. 4. TGA profile of film TGG02. measured during the third cycle of the measurement procedure. An
From the thermogram presented in Fig. 4, two weight losses are example of the glass transition region is presented in Fig. 5, in which the
clearly detected. Decomposition of fish gelatin-glycerol polymeric DSC curve of the TGG02 film is showed, in which the Tg is appears
structure is detected around 300 ◦ C, whereas Tara powder degradation around 62 ◦ C. The DSC curves obtained from all the films, in which the
begins at 550 ◦ C. The complete set of thermograms obtained from all the Tg region is observed, is shown in Fig. S7 of the ESI.
films fabricated are presented in Fig. S6 of the ESI.

5
L. Nuvoli et al. Polymer xxx (xxxx) xxx

Table 2 [31]. Finally, the humidity content values for TGG films vary between
Thermal properties of all the films prepared. Tg is the glass transition tempera­ the limit values marked for pure Fish gelatin film (4.1%) and pure Tara
ture, humidity loss percentage was measured directly as the quantity of mass loss film (11.7%). In this case, the use of milled Tara reduces the humidity
at 100 ◦ C, T5% and T10% are the temperatures at which 5% and 10% of mass is content respect to films with non-milled Tara (compare, for example,
lost, and Onset temperature is defined as the temperature at which the decom­ humidity content values of TGG 01 and TGG 02 films). This is also a very
position of the material begins.
positive effect in terms of stability and handleability of the films. It is
Film Tg (◦ C) Humidity loss (%) T5% (◦ C) T10% (◦ C) Onset (◦ C) important to remark that similar humidity loss and thermal decompo­
TGG 01 63 9.8 213 245 274 sition profiles were reported by Pulieri et al. in the case of Chito­
TGG 02 62 6.9 224 268 294 san/gelatin blends [32].
TGG 03 52 10.7 190 218 266 Concerning the analysis of the glass transition temperature, it
TGG 04 54 10.1 193 237 287
emerges that TGG films show higher Tg values with respect to FG/Gly,
TGG 05 50 10.5 175 207 259
TGG 06 51 7.3 244 261 275 although this difference is not very important (in Ta/Gly film was not
TGG 07 69 9.7 207 244 274 possible to measure the Tg temperature precisely). The discussion of the
TGG 08 61 8.1 215 251 277 results can be carried out taking into account three different separated
TGG 09 63 6.1 217 263 287
effects: (i) The influence of the Ta/FG ratio on the Tg, (ii) The effect of
TGG 10 58 5.3 228 268 293
FG/Gly 48 4.1 121 191 264 glycerol weight percentage, and (iii), the type of Ta employed (milled or
Ta/Gly 52 11.7 258 269 319 not milled).
Fig. 6 shows the Tg variation in all the films as a function of the Ta/FG
ratio (Fig. 5a) and also as a function of glycerol weight percentage
All the thermal data derived from TGA and DSC tests corresponding (Fig. 5b).
to all the films fabricated are reported in Table 2.
As it can be seen in data in Table 2, pure FG films (FG/Gly) present
the lower T5%, T10% and onset temperature values, with temperatures of Table 3
121 ◦ C, 191 ◦ C and 264 ◦ C, respectively. Comparing these data with the Mechanical data of the films TGG01-TGG10.
thermal behaviour of pure Tara films (Ta/Gly), values are considerably Film E Δa σ break Δa ε break Δa
increased (T5%, T10% and onset values are 258 ◦ C, 269 ◦ C and 319 ◦ C), (MPa) (MPa) (MPa) (MPa) (%) (%)

indicating that the Tara powder has a better thermal stability that Fish TGG 97 13.3 24 2.5 75 7.3
gelatin. In both cases, the quantity of glycerol employed is 20 wt%. 01
TGG 195 18.8 23 1.5 58 6.4
Values of T5%, T10% and onset temperature in Tara-gelatin films
02
(TGG01 to TGG10) lie between the limit values showed by the FG/Gly TGG 11 2.7 9 2.9 100 11.1
and Ta/Gly. In this sense, it is demonstrated the good thermal behaviour 03
of composite films respect to pure FG film. This improvement on the TGG 27 5.4 15 3.1 73 9.2
thermal behaviour depends strongly on the quantity of glycerol, but we 04
TGG 5 1.2 4 0.3 107 8.6
can even remark that films with high glycerol content (TGG 05 and TGG
05
06, with 60 wt% of glycerol) present a good thermal stability (onset TGG 12 4.4 7 2.5 96 14.1
temperatures are 259 ◦ C and 275 ◦ C). 06
As expected, the quantity of glycerol has a predominant effect in the TGG 194 19.8 27 3.1 62 13.2
07
thermal stability of the films. In this sense, increasing the glycerol
TGG 94 21.1 13 2.1 78 4.1
content lowers the T5% and T10% temperatures, then affecting negatively 08
to the thermal stability and to the processing or working temperatures. TGG 234 17.8 35 5.2 74 17.2
On the other hand, using milled Tara seems to affect positively in the 09
thermal stability of the films. For example, comparing the TGA data of TGG 200 21.2 23 1.5 63 12.3
10
TGG 01 (non-milled Tara) and TGG 02 (milled Tara), it can be seen that
FG/Gly 243 5.9 36 3.8 95 6.1
T5%, T10% and onset temperatures are increased up to 20 ◦ C, indicating Ta/Gly 550 31.7 40 13.2 23 7.0
that the use of milled Tara also increases the thermal stability of the a
Δ indicate the standard deviation of each value.
films, obtaining films stable up to 300 ◦ C, making them easy processable

Fig. 6. Tg values of films fabricated. a) Films with different Ta/FG ratio; b) Films with different glycerol weight percentage.

6
L. Nuvoli et al. Polymer xxx (xxxx) xxx

Fig. 7. Dependence of the mechanical data of the films with glycerol content. a) Young’s moduli, b) Stress at break, and c) Deformation at break.

Fig. 6a shows the influence of the Ta/FG ratio in the glass transition
temperature. Having a look at films fabricated using non milled Tara, a
reduction of Tg is observed when the Ta/FG ratio increases. On the other
hand, the effect of the glycerol weight percentage is clearly observed
(Fig. 6b). As expected, the addition of glycerol decreases the Tg, con­
firming the role of glycerol as plasticizer. Finally, the third effect is
related to the Tara powder employed (non-milled or milled). The anal­
ysis of the data in Fig. 6 shows that using milled Tara increases the glass
transition temperature, but only when TGG07 vs TGG08 and TGG09 vs
TGG10 films are compared (see Fig. 6a). In these particular cases, Tg is
increased around 7 ◦ C.

5.6. Tensile properties

To analyse the mechanical behaviour of films TGG01-TGG10, tensile

properties were measured. Three different parameters were determined
from the stress (σ)-strain (ε) curves: Young modulus E, stress (σ break,
(MPa)) and deformation at break (ε break, (%)). Fig. S8 of the ESI shows
the stress-strain curves obtained for all the films fabricated, while
Table 3 presents the corresponding mechanical data.
Mechanical data presented in Table 3 indicate that Young’s moduli Fig. 8. Variation of Young modulus E with the Ta/FG ratio and effect of the
values show a wide variability, from 5 MPa (TGG 05) to 234 MPa (TGG ball-milling of Tara gum powder.
09), as well as the stress at break, included between 4 MPa (TGG05) and
35 MPa (TGG09). Consequently, the deformation at break range be­ content is reported in Fig. 7.
tween 23% and 107%. It can be observed that the Young’s moduli, as well as the stress at
This variability in the mechanical data can be related to the glycerol break decrease when glycerol is added, while the deformation at break
content and its plasticization effect, which plays a key role in the me­ increases with the content of glycerol. This behaviour confirms the
chanical performance of the materials. In order to better highlight this plasticization effect expected through the addition of glycerol in the
effect, the dependence of the mechanical parameters with the glycerol initial formulation of the materials.

7
L. Nuvoli et al. Polymer xxx (xxxx) xxx

Fig. 9. Mechanical data as function of Ta/FG of films containing non-milled or milled Tara gum powder. a) Stress at break; b) Deformation at break.

Having a look at the values individually, TGG09 and TGG10 films (Tara in our case) represents a novelty and can lead the way toward
present the higher values of Young’s moduli (234 and 200 MPa). This is further optimization of similar polymers.
directly related to the Tara powder content, which is higher in these
films (2 wt eq). Data availability
In addition to the two main effects discussed above, a not-negligible
influence of the Tara grain size on the mechanical properties can be The raw data required to reproduce these findings cannot be shared
observed. In Fig. 8, the variation of E as function of Ta/FG ratio and type at this time due to technical limitations.
of Tara (milled and not milled) is reported.
From the plots showed in Fig. 8 it is possible to notice a different
mechanical behaviour of films containing milled Ta which influences Declaration of competing interest
differently the trends of the two series TGG07-TGG01-TGG09 and
TGG08-TGG02-TGG10 (Fig. 8, circled and squared points). Films pre­ The authors declare that they have no known competing financial
pared with not milled Tara show a difference in Young’s moduli value of interests or personal relationships that could have appeared to influence
about 100 MPa for Ta/FG ½ ratio (TGG07 vs TGG08) and for Ta/FG ratio the work reported in this paper.
of 1/1 (TGG02 vs TGG01), but with opposite trends. In the presence of
an excess of Tara, the Young’s moduli values are quite similar, with not Acknowledgments
milled films showing a value slightly higher. Finally, to better compare
the curves relative to films containing non-milled and milled Tara, the The financial support provided by FEDER (Fondo Europeo de
variation of stress at break and deformation at break of films containing Desarrollo Regional) and both the Spanish Agencia Estatal de Inves­
the two different Tara powders is reported in Fig. 9. tigacióon (MAT2017–84501-R; MAT2017-88923-P) and the Consejería
Data in Fig. 9 indicates that stress at brake is affected by the presence de Educación, Junta de Castilla y León (BU306P18) is gratefully
of milled Tara only in films containing with Ta/FG ratio 2/1 or ½, while acknowledged.
the mechanical values for Ta/FG ratio of 1/1 are almost coincident
(Fig. 9a). On the contrary, the deformation at brake shows a different Appendix A. Supplementary data
behaviour depending of the Tara employed. When non-milled Tara is
used, deformation at break grows with the increasing of Tara content, Supplementary data to this article can be found online at https://doi.
whereas on the other hand when milled Tara is employed the trend is the org/10.1016/j.polymer.2020.123244.
opposite (Fig. 9b).
References
6. Conclusions
[1] R. Rodríguez-Rodríguez, H. Espinosa-Andrews, C. Velasquillo-Martínez, Z.
Y. García-Carvajal, Composite hydrogels based on gelatin, chitosan and polyvinyl
Morphological, thermal and mechanical properties of films
alcohol to biomedical applications: a review, Int. J. Polym. Mater. 69 (2015) 1–20,
composed by fish gelatin, Tara gum and glycerol can be tuned by https://doi.org/10.1080/00914037.2019.1581780.
oportune engineering processes. In particular, by changing the amount [2] T. Huang, Z. Fang, H. Zhao, D. Xu, W. Yang, W. Yu, J. Zhang, Physical properties
of the plasticizer glycerol, it is possible to optimize the composition in and release kinetics of electron beam irradiated fish gelatin films with antioxidants
of bamboo leaves, Food Bioscience, 36 100597 (2020), https://doi.org/10.1016/j.
terms of thermal behaviour and mechanical properties. Also, the struc­ fbio.2020.100597.
tural effect obtained by the addition of Tara gum resulted relevant. By [3] T. Huang, J. Lin, Z. Fang, W. Yu, Z. Li, D. Xu, W. Yang, J. Zhang, Food and, Bioproc.
employing milled Tara instead to the commercial one, it is possible to Technol. 13 (2020) 522–532, https://doi.org/10.1007/s11947-020-02409-w.
[4] P. Díaz-Calderón, L. Caballero, F. Melo, J. Enrione, Molecular configuration of
enhance the thermal stability and improve the mechanical performances gelatin–water suspensions at low concentration, Food Hydrocolloids 39 (2014)
of the films. The origin of such effects can be found in a change of the 171–179, https://doi.org/10.1016/j.foodhyd.2013.12.019.
morfology of films prepared with different ingredients, and observed by [5] Q. Xing, K. Yates, C. Vogt, Z. Qian, M.C. Frost, F. Zhao, Increasing mechanical
strength of gelatin hydrogels by divalent metal ion removal, Sci. Rep. 4 (2014)
IR spectroscopy and through SEM and AFM analyses. Influencing the 1–10, https://doi.org/10.1038/srep04706.
performences of the films by reducing the grain size of the natural gum [6] H. Massoumi, J. Nourmohammadi, M.S. Marvi, F. Moztarzadeh, Comparative study
of the properties of sericin-gelatin nanofibrous wound dressing containing

8
L. Nuvoli et al. Polymer xxx (xxxx) xxx

halloysite nanotubes loaded with zinc and copper ions, Int. J. Polym. Mater. Polym. composite gels, J. Food Sci. Technol. 54 (2017) 518–530, https://doi.org/
Biomater 68 (2019) 1–12, https://doi.org/10.1080/00914037.2018.1534115. 10.1007/s13197-017-2496-9.
[7] W.W. Thein-Han, J. Saikhun, C. Pholpramoo, R.D.K. Misra, Y. Kitiyanant, [20] Y. Wu, W. Ding, L. Jia, Q. He, The rheological properties of tara gum (Caesalpinia
Chitosan–gelatin scaffolds for tissue engineering: physico-chemical properties and spinosa),, Food Chem. 168 (2015) 366–371, https://doi.org/10.1016/j.
biological response of buffalo embryonic stem cells and transfectant of GFP–buffalo foodchem.2014.07.083.
embryonic stem cells, Acta Biomater. 5 (2009) 3453–3466, https://doi.org/ [21] A. Mortensen, F. Aguilar, R. Crebelli, A. Di Domenico, M.J. Frutos, P. Galtier,
10.1016/j.actbio.2009.05.012. D. Gott, U. Gundert-Remy, C. Lambré, J.-C. Leblanc, O. Lindtner, P. Moldeus,
[8] X. Feng, V.K. Ng, M. Mikš-Krajnik, H. Yang, Effects of fish gelatin and tea P. Mosesso, A. Oskarsson, D. Parent-Massin, I. Stankovic, I. Waalkens-Berendsen, R.
polyphenol coating on the spoilage and degradation of myofibril in fish fillet A. Woutersen, M. Wright, M. Younes, L. Brimer, A. Christodoulidou, F. Lodi,
during cold storage, Food Bioprocess Technol. 10 (2017) 89–102, https://doi.org/ A. Tard, B. Dusemund, Re-evaluation of tara gum (E 417) as a food additive, EFSA
10.1007/s11947-016-1798-7. Journal 15 (6) (2017) e04863, https://doi.org/10.2903/j.efsa.2017.4863.
[9] J. Rose, S. Pacelli, A. Haj, H. Dua, A. Hopkinson, L. White, F. Rose, Gelatin-based [22] http://www.lapigelatine.com/wp-content/uploads/2017/01/fish.gelatine.edible.
materials in ocular tissue engineering, Materials 7 (7) (2014) 3106–3135, https:// grade_.pdf (accessed on 10/10/2020).
doi.org/10.3390/ma7043106. [23] J. Chong, O. Soufan, C. Li, I. Caraus, S. Li, G. Bourque, D.S. Wishart, J. Xia,
[10] J.R. Dias, S. Baptista-Silva, C.M. Oliveira, A. de Sousa, A.L. Oliveira, P.J. Bartolo, P. MetaboAnalyst 4.0: towards more transparent and integrative metabolomics
L. Granja, In-situ crosslinked electrospun gelatin nanofibers for skin regeneration, analysis, Nucleic Acids Res. 46 (2018) 486–494, https://doi.org/10.1093/nar/
Eur. Polym. J. 95 (2017) 161–173, https://doi.org/10.1016/j. gky310.
eurpolymj.2017.08.015. [24] F. Delogu, G. Gorrasi, A. Sorrentino, Fabrication of polymer nanocomposites via
[11] M.S. Rahman, G.S. Al-Saidi, N. Guizani, Thermal characterisation of gelatin ball milling: present status and future perspectives, Prog. Mater. Sci. 86 (2017)
extracted from yellowfin tuna skin and commercial mammalian gelatin, Food 75–126, https://doi.org/10.1016/j.pmatsci.2017.01.003.
Chem. 108 (2008) 472–481, https://doi.org/10.1016/j.foodchem.2007.10.079. [25] B.S. Pascual, M. Trigo-López, J.A. Reglero Ruiz, J.L. Pablos, J.C. Bertolín,
[12] A. Duconseille, T. Astruc, N. Quintana, F. Meersman, V. Sante-Lhoutellier, Gelatin C. Represa, J.V. Cuevas, Félix C. García, J. M. García, Porous aromatic polyamides
structure and composition linked to hard capsule dissolution: a review,, Food the easy and green way, European Polymer Journal 116 (2019) 91–98, https://doi.
Hydrocolloids 43 (2015) 360–376, https://doi.org/10.1016/j. org/10.1016/j.eurpolymj.2019.03.058.
foodhyd.2014.06.006. [26] S. Basu, U.S. Shivhare, T.V. Singh, V.S. Beniwal, Rheological, texture and spectral
[13] E. Jeevithan, Z. Qingbo, B. Bao, W. Wu, Biomedical and pharmaceutical characteristics of sorbitol substituted mango jam, J. Food Eng. 105 (2011)
application of fish collagen and gelatin: a review,, J. Nutr. Therapeut. 2 (2013) 503–512, https://doi.org/10.1016/j.jfoodeng.2011.03.014.
218–227. [27] UNE-EN 13432, Requirements for packaging recoverable through composting and
[14] A. Etxabide, I. Leceta, S. Cabezudo, P. Guerrero, K. de la Caba, Sustainable fish biodegradation, Test scheme and evaluation criteria for the final acceptance of
gelatin films: from food processing waste to compost, ACS Sustain. Chem. Eng. 4 packaging (2001) 2001.
(2016) 4626–4634, https://doi.org/10.1021/acssuschemeng.6b00750. [28] J.M. Chalmers, N.J. Everall, Polymer analysis and characterization by FTIR, FTIR-
[15] A. Mannu, S. Garroni, J. Ibanez Porras, A. Mele, Available technologies and microscopy, Raman spectroscopy and chemometrics, Int. J. Polym. Anal. Char. 5
materials for waste cooking oil recycling, Processes 8 (2020) 366, https://doi.org/ (1999) 223–245, https://doi.org/10.1080/10236669908009739.
10.3390/pr8030366. [29] M.S. Lindblad, B.M. Keyes, L.M. Gedvilas, T.G. Rials, S.S. Kelley, FTIR imaging
[16] M. Bocqué, C. Voirin, V. Lapinte, S. Caillol, J.J. Robin, Petrobased and bio-based coupled with multivariate analysis for study of initial diffusion of different solvents
plasticizers: chemical structures to plasticizing properties, J. Polym. Sci., Part A: in cellulose acetate butyrate films, Cellulose 15 (2008) 23–33, https://doi.org/
Polym. Chem. 54 (2016) 11–33, https://doi.org/10.1002/pola.27917. 10.1007/s10570-007-9173-5.
[17] A.A. Karim, R. Bhat, Fish gelatin: properties, challenges, and prospects as an [30] B. Worley, R. Powers, Multivariate analysis in metabolomics, Curr. Metabolomics 1
alternative to mammalian gelatins, Food Hydrocolloids 23 (2009) 563–579, (2013) 92–107, https://doi.org/10.2174/2213235X11301010092.
https://doi.org/10.1016/j.foodhyd.2008.07.002. [31] M.A. Meador, Recent advances in the development of processable high-
[18] M. Ramos, A. Valdés, A. Beltrán, M.C. Garrigós, Gelatin-based films and coatings temperature polymers, Annu. Rev. Mater. Sci. 28 (1998) 599–630, https://doi.org/
for food packaging applications, Coatings 6 (2016) 41, https://doi.org/10.3390/ 10.1146/annurev.matsci.28.1.599.
coatings6040041. [32] E. Pulieri, V. Chiono, G. Ciardelli, G. Vozzi, A. Ahluwalia, C. Domenici, F. Vozzi,
[19] P.K. Binsi, N. Nayak, P.C. Sarkar, C.G. Joshy, G. Ninan, C.N. Ravishankar, Gelation P. Giusti, Chitosan/gelatin blends for biomedical applications, J. Biomed. Mater.
and thermal characteristics of microwave extracted fish gelatin-natural gum Res. 86 (2008) 311–322, https://doi.org/10.1002/jbm.a.31492.