Food Hydrocolloids 97 (2019) 105173

Contents lists available at ScienceDirect

Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Lime peel pectin integrated with coconut water and lime peel extract as a T
new bioactive film sachet to retard soybean oil oxidation
Pattrathip Rodsamrana,c, Rungsinee Sothornvita,b,∗
a
Department of Food Engineering, Faculty of Engineering at Kamphaengsaen, Kasetsart University, Kamphaengsaen Campus, Nakhonpathom, 73140, Thailand
b
Center of Advanced Studies in Industrial Technology, Kasetsart University, Bangkok, 10900, Thailand
c
School of Culinary Arts, Suan Dusit University, Bangkok, 10700, Thailand

A R T I C LE I N FO A B S T R A C T

Keywords: Lime peel pectin films incorporated with natural antioxidants from lime peel residue extract and using coconut
Pectin film water (CW) as a plasticizer were developed and applied as a sachet to retard the oxidation of soybean oil. The
Lime peel films prepared with CW as plasticizer had the lowest water solubility and were flexible compared to films
Antioxidant prepared with glycerol as plasticizer. High water vapor permeability was observed in the films with higher
Soybean oil
plasticizer amounts in both CW and glycerol. However, the combination of CW and lime peel extract improved
Coconut water
the film water barrier. The incorporation of lime peel extract in the film increased the total phenolic content and
therefore enhanced the antioxidant activity of the film. Moreover, all lime peel pectin films retarded soybean oil
oxidation during 30 days of storage. These results indicated that lime peel residue was a good source for pectin
production and a natural antioxidant in bioactive films or coatings for various food applications.

1. Introduction montmorillonite. Moreover, Bernhardt, Pérez, Fissore, De’Nobili, and

Bio-based films and coatings have been extensively developed in the
last few decades. Most biodegradable or edible materials are based on
polysaccharides, proteins and lipids. Pectin is one of the versatile
polysaccharides considered an effective biomaterial film due to its
biodegradability, biocompatibility and non-toxicity (Nisar et al., 2018).
Pectin is extracted from by-products of juice manufacturing and
other agricultural wastes such as lime (Fishman, Chau, Hoagland, &
Hotchkiss, 2006), passion fruit peel (Seixas et al., 2014) and pome-
granate peel (Abid et al., 2016, 2017). From previous study, lime peel
pectin was extracted using microwave heating with a short processing
time and its properties were characterized as a high methoxyl pectin
having rapid-set gel formation (Rodsamran & Sothornvit, 2019b).
Pectin films act as a good oxygen barrier and have good hardness and
adhesiveness (Bermúdez-Oria, Rodríguez-Gutiérrez, Vioque, Rubio-
Senent, & Fernández-Bolaños, 2017). However, some limitations need
to be solved, such as brittleness, rigidity, high water sensitivity and
poor water barrier of pectin films (Farris et al., 2009). A blend of pectin
and other ingredients may improve the pectin-based film properties and
thus be more suitable for food application. Oliveira et al. (2016) im-
proved the water vapor permeability, tensile strength and elastic
modulus of pomegranate peel pectin films by adding 6%
Rojas (2017) found that 5% of corn husk fiber filler improved the
performance regarding water vapor permeability and tensile strength in
low methoxyl pectin films. Besides, the fruit purees (mango or guava)
produced with pectin film was improved by addition of nanofibrillated
bacterial cellulose contents to replace pectin resulting in stronger films
with stiffness, more water resistance and water barrier (Viana, Sa,
Barros, Borges, & Azeredo, 2018). They suggested to apply fruit purees
based pectin films for sachet application. Furthermore, the pectin films
incorporated with spent coffee grounds increased water vapor perme-
ability and thermal stability (Mendes et al., 2019). In addition, edible
films based on pectin have been incorporated with several active
compounds, including antioxidants, antimicrobials, antibrowning
agents and flavoring (Espitia, Du, de Jesús Avena-Bustillos, de Fátima
Ferreira Soares, & McHugh, 2014; Falguera, Quintero, Jiménez, Muñoz,
& Ibarz, 2011).
Nowadays, agricultural by-products are considered as a source of
active ingredients like antioxidants and antimicrobials (Galanakis,
2013). Lime peel was extracted for total phenolic compounds using
ultrasonic-assisted extraction and showed a high antioxidant activity
with a saving in time compared with microwave-assisted extraction
(Rodsamran & Sothornvit, 2019a). Thus, the addition of natural extract
into bio-based films has been considered for improving film properties.

∗
Corresponding author. Department of Food Engineering, Faculty of Engineering at Kamphaengsaen, Kasetsart University, Kamphaengsaen Campus,
Nakhonpathom, 73140, Thailand.
E-mail address: fengrns@ku.ac.th (R. Sothornvit).

https://doi.org/10.1016/j.foodhyd.2019.105173
Received 14 April 2019; Received in revised form 12 June 2019; Accepted 18 June 2019
Available online 19 June 2019
0268-005X/ © 2019 Elsevier Ltd. All rights reserved.
P. Rodsamran and R. Sothornvit Food Hydrocolloids 97 (2019) 105173

For example, the incorporation of hydroxytyrosol and 3,4-dihydrox- Table 1

yphenylglycol from olive fruit in pectin-fish gelatin films exhibited Formulations of the lime peel pectin films.
good barrier properties and also preserved strawberries against mold Sample Lime peel Water (g) Coconut Glycerol (g) Lime peel
during storage (Bermúdez-Oria et al., 2017). Nisar et al. (2018) re- pectin (g) water (g) extract (g)
ported that the inclusion of clove bud essential oil in citrus pectin films
G20 1.0 99.0 0.0 0.2 0.0
enhanced the water barrier properties and resistant to breakage, im-
G30 1.0 99.0 0.0 0.3 0.0
proved flexibility and increased the total phenolic content and radical CW10 1.0 89.1 9.9 0.0 0.0
scavenging activity. Furthermore, Rodsamran and Sothornvit (2018) CW20 1.0 79.2 19.8 0.0 0.0
found that coconut protein films using mature coconut water as a sol- G20E 1.0 99.0 0.0 0.2 20.0
vent and incorporated with coconut water concentrate extracted using G30E 1.0 99.0 0.0 0.3 20.0
CW10E 1.0 89.1 9.9 0.0 20.0
evaporation, improved the mechanical properties of films and increased
CW20E 1.0 79.2 19.8 0.0 20.0
the total phenolic content and antioxidant capacity. Dos Santos Caetano
et al. (2018) showed that the addition of minimally processed pumpkin
residue extract and oregano essential oil to cassava starch films im- 2.3. Natural extract from lime peel extraction
proved the mechanical and antioxidant properties of the films. More-
over, apple peel polyphenol extract incorporated into chitosan films The antioxidant was extracted from the lime peel extract according
provided novel functional films with good mechanical and anti- to the method of Rodsamran and Sothornvit (2019a). Briefly, lime peel
microbial properties (Riaz et al., 2018). Additionally, bioactive edible powder (1.5 ± 0.005 g) and 30 mL of 55% ethanol were vortexed for
packaging films incorporated with tea polyphenol into pomelo peel 30 s. An ultrasound instrument (VCX 750, Sonics & Materials, Inc.,
flours significantly improved antioxidant activity and antimicrobial Newtown, CT, USA) with a stepped microtip size of 3 mm was used for
activity and the bioactive films decreased peroxide value of soybean oil extraction at 38% amplitude for 4 min. After cooling for 30 s, the nat-
indicating the reduction of oil oxidation during storage at 50 °C for 30 ural extract was filtered through Whatman No. 93 filter paper and the
days (Wu et al., 2019). volume was adjusted to 30 mL using 55% ethanol. The natural extract
To date, there has been no report on the use of antioxidants from from the lime peel was collected at −18 °C in an amber glass bottle.
mature coconut water and lime peel extract to form new bioactive films
based on lime peel pectin. Therefore, this study examined the physi- 2.4. Lime peel pectin film preparation
cochemical, water barrier, mechanical and antioxidant properties of
lime peel pectin films integrated with mature coconut water and nat- Film-forming solution (1% w/w) was prepared by moistening lime
ural extract from lime peel. Additionally, the antioxidant effect of the peel pectin with a small amount of ethanol and dissolving in water
film, with and without the antioxidant extract, was evaluated on soy- using two types of plasticizer (glycerol and coconut water) at two levels
bean oil oxidation. of concentrations (Table 1) for 2 h at 85 °C with magnetic stirring. After
cooling to room temperature, the natural extract from lime peel was
2. Materials and methods added into the respective film solution at 20% (w/w) of the film solu-
tion. Then, the film forming solutions were left to stand at 4 °C for 6–8 h
2.1. Materials to remove air bubbles. Later, they were cast onto sterile plastic Petri
dishes (diameter 15 cm) and dried at 45 °C for 18 h to obtain
Lime peel residue, provided by beverage shops on the Kasetsart 0.9 ± 0.1 g of dried films. Finally, the obtained lime peel pectin films
University Kamphangsaen campus (Nakhonpathom, Thailand), was cut were peeled off and conditioned at 25 ± 3 °C and 50 ± 5% RH for
and dried at 60 °C for 12 h using a tray dryer (Mallory Quality Product, 48 h prior to testing.
Bangkok, Thailand). The lime peel powder was obtained by milling
through a 1 mm screen in a hammer rotor grinding mill (Retsch GmbH, 2.5. Film characterizations
Haan, Germany) and packing in vacuum sealed plastic bag at room
temperature (28 ± 2 °C). Mature coconut water (CW) was donated by 2.5.1. Thickness, moisture content and water solubility
Theppadungporn Coconut Co., Ltd. (Nakhonpathom, Thailand) and Film thickness was measured with a micrometer (No. 7326;
sterilized at 121 °C for 5 min in a 200 mL glass bottle using an autoclave Mitutoyo Manufacturing Co., Ltd, Tokyo, Japan) at five random posi-
(Hirayamaha–300 MIT, Tokyo, Japan). All other reagents were supplied tions around the film. An average of film thickness for each film re-
by U&V Holding (Thailand) Co., Ltd. (Nonthaburi, Thailand). plicate was used to calculate the mechanical and water barrier prop-
erties.
Film samples (20 mm × 20 mm) were weighed (W1) and dried in a
2.2. Lime peel pectin extraction hot-air oven at 105 °C until constant weight (W2). Then the film pieces
were immersed into 20 mL of distilled water and shaken at 25 ± 0.1 °C
The lime peel pectin extraction was modified from Rodsamran and for 24 h. The residual films were separated and dried at 105 °C for 24 h
Sothornvit (2019b). The lime peel powder (50 g) was mixed in 2 kg of until constant weight (W3). The moisture content and water solubility
0.05M citric acid and preheated in a microwave oven (White Wes- were calculated according to the following equations:
tinghouse, KM90VP–63/03, Pittsburg, PA, USA) with an output power
of 1400 W for 10 min to reach 85–90 °C. The pectin extract solution was W1 − W2
Moisture (%) = × 100
obtained by holding the mixture at a microwave output power of 420 W W1 (1)
for 30 min. After filtration through a 420 μm screen, the filtrate was W2 − W3
centrifuged at 10,000✕g (25 °C) for 5 min and the clear pectin solution Solubility (%) = × 100
W2 (2)
was collected and purified using precipitation with double the volume
of absolute ethanol. Lime peel pectin was separated from the liquid,
washed three times with absolute ethanol, and given a final washing 2.5.2. Opacity
with acetone. After the acetone odor had disappeared, the purified The opacity of films was determined using a UV–visible spectro-
pectin was dried at 50 °C for 1 h using a hot air oven (Redline RF 115, photometer (V-770 UV/VIS/NIR, Jasco, Tokyo, Japan) according to the
Tuttlingen, Germany), ground in a Waring blender (HGB2WT, Torring, method proposed by Rodsamran and Sothornvit (2018). Rectangular
CT, USA), and sieved using a 125 μm screen. film strips (10 mm × 40 mm) were directly placed in a

2
P. Rodsamran and R. Sothornvit Food Hydrocolloids 97 (2019) 105173

Table 2
Physico-chemical, water vapor permeability (WVP) and mechanical properties of lime peel pectin films.
Film property Film sample∗

G20 G30 CW10 CW20 G20E G30E CW10E CW20E

c a bc bc ab ab ab
Moisture (%db) 11.90 ± 2.26 25.62 ± 6.09 18.16 ± 0.34 18.20 ± 1.87 20.72 ± 0.12 21.92 ± 3.25 19.59 ± 0.18 25.67a ± 0.62
Solubility (%) 43.98b ± 2.09 49.43b ± 1.97 27.26d ± 3.48 36.61c ± 2.88 62.43a ± 2.23 3.35a ± 3.50 64.04a ± 0.40 67.14a ± 1.76
Opacity (Abs600⋅ 2.62de ± 0.64 2.27e ± 0.10 2.41e ± 0.11 3.85bc ± 0.44 3.39cd ± 0.34 5.56a ± 0.20 4.39b ± 0.58 3.79bc ± 0.31
mm−1)
Thickness (μm) 48.27a ± 0.85 48.48a ± 2.36 48.62a ± 1.31 49.77a ± 0.75 48.62a ± 2.09 48.28a ± 2.48 48.58a ± 1.74 49.83a ± 0.76
WVP (✕10−10 g ⋅ 1.86c ± 0.12 2.06bc ± 0.08 1.89c ± 0.07 2.55a ± 0.04 2.25b ± 0.11 2.12b ± 0.04 1.63d ± 0.09 2.12b ± 0.11
m−1s−1Pa−1)
Tensile strength 16.93b ± 2.44 13.49c ± 0.77 20.61a ± 3.69 5.62ef ± 0.63 8.57de ± 1.09 14.00bc ± 1.82 11.06cd ± 0.42 3.63f ± 0.44
(MPa)
Elastic modulus 947.70ab ± 20.35 577.44c ± 20.62 974.77a ± 60.73 227.11e ± 15.67 904.45b ± 18.20 441.08d ± 43.17 463.45d ± 33.21 90.95f ± 1.62
(MPa)
Elongation (%) 1.77f ± 0.10 3.11cd ± 0.20 2.31def ± 0.31 3.77bc ± 0.20 2.11ef ± 0.20 4.60ab ± 1.17 2.78de ± 0.38 4.78a ± 0.70

Values are expressed as mean of two replicates ± standard deviation.

Different lowercase superscript letters in the same row indicate significantly different values (p < 0.05).
∗
The abbreviations of film samples are following; G means glycerol, CW means coconut water, E means lime peel extract and the numbers following the letters mean
the amount used. For example, G20, CW10 and CW10E represent films plasticized with 20% glycerol, 10% coconut water and 10% coconut water incorporated with
lime peel extract, respectively.

spectrophotometer test cell and absorbance was read at a wavelength of antioxidant activity was calculated using the standard curve of Trolox
600 nm. The film opacity was calculated by dividing the absorbance at solution and expressed as micromoles Trolox equivalents per gram of
600 nm by the thickness of the film strip (A600⋅mm−1). dried film sample (μM Trolox/g film).

2.5.3. Water vapor permeability
The water vapor permeability (WVP) of the pectin films was mea-
sured following the gravimetric modified cup method based on ASTM
E96 (McHugh, Avena-Bustillas, & Krochta, 1993). Cups were filled with
7 mL distilled water and sealed tightly with film specimens, then placed
at 25 ± 3 °C (50 ± 5% RH) in a cabinet. The cups were weighed
periodically for 12 h. The water vapor transmission rate (WVTR) was
obtained by dividing the slope of the regression analysis of weight loss
data versus time by the film area. WVP values were calculated by
multiplying the WVTR by the film thickness and dividing by the water
vapor partial pressure difference across the films.
2.5.4. Mechanical properties
Tensile strength, elastic modulus and percentage of elongation at
break of the pectin films were measured using a universal testing ma-
chine (Model 5569, Instron, Norwood, MA, USA) with a 50 N load cell,
10 mm/min crosshead speed and 50 mm initial grip separation. Film
specimens of 8 mm × 50 mm were tested at 23 ± 2 °C and 50 ± 5%
RH according to ASTM D882–97 (ASTM, 1997).
2.5.5. Total phenolic content
The film extract solutions were prepared by dissolving 300 mg of
film specimen in 30 mL of water or ethanol, homogenizing at 5000 rpm
for 30 s, then centrifuging at 10,000×g for 10 min at 25 °C. The clear
solutions were collected and their total phenolic contents (TPC) were
determined using the modified method of Srivastava et al. (2010). For
this purpose, 0.25 mL of each film extract solution was first reacted
with 0.25 mL of 10% Folin-Ciocalteu solution for 5 min, and then
0.5 mL of 7.5% (w/w) of sodium carbonate and 4 mL of distilled water
were added. After that, the solution was mixed thoroughly and kept in a
dark chamber at room temperature for 1 h. The absorbance was mea-
sured at 765 nm and TPC was expressed as milligrams gallic acid
equivalents per gram of dried film sample (mg GAE/g film).
2.5.6. Antioxidant activities
The film extract solutions obtained from TPC determination were
used to evaluate the antioxidant activity of the film. The 2,2′-diphenyl-
1-picrylhydrazyl (DPPH) assay and 2,2′-azino-bis(3-ethylbenzothiazo-
line-6-sulfonic acid) diammonium salt (ABTS) assay were estimated
according to the method of Rodsamran and Sothornvit (2018). The
2.6. Thiobarbituric acid reactive substances (TBARs) determination
Soybean oil (3 mL) was added into polypropylene film (PP-film) and
the selected pectin film bag (30 mm × 40 mm) and sealed using a hand-
pressed desktop sealer (PFS-2000, PowerGoldex, Zhejiang, China). The
samples were kept at 27 ± 2 °C (50 ± 5% RH) under fluorescent light
and taken after 0, 2, 4, 6, 10, 14, 20 and 30 days for the determination
of oil oxidation according to the method of Benjakul, Visessanguan,
Phongkanpai, and Tanaka (2005) with some modification. The oil
(0.5000 ± 0.05 g) was mixed with 5 mL of a solution containing
0.0375% thiobarbituric acid, 15% trichloroacetic acid and 0.875 mL
hydrochloric acid. The mixture was heated in a water bath at 95 °C for
20 min, followed by cooling to room temperature and centrifuging at
12,000×g for 5 min. The absorbance of supernatant was read at
532 nm. TBARs was calculated from the standard curve of mal-
onaldehyde (MDA) and expressed as milligrams MDA per kilogram of
oil (mg MDA/kg oil). Soybean oil without the film covering was used as
the control.
2.7. Statistical analysis
The results of film characterization were subjected to one-way
analysis of variance with a completely randomized experimental de-
sign. Means were tested using Duncan's multiple range tests at a sig-
nificance level of 0.05, with the SPSS software (Version 11.5, SPSS®
Inc., Chicago, IL, USA). All experiments were duplicated.
3. Results and discussion
3.1. Thickness, moisture content and water solubility
There was no significant difference on the thickness of all lime peel
pectin based films. The average film thickness was 48.73 ± 1.75 μm
(Table 2). The total solid of dried films was controlled in order to
compare all film properties. Generally, the thickness of pectin films was
thin due to the compact molecular structure. Similar result was found in
the pectin films from the work of Mendes et al. (2019). As expected, the
films with higher concentrations of glycerol (G30) exhibited higher
moisture contents (Table 2). An increase in glycerol concentration led
to an increase in molecular mobility. Similar results were reported for

3
P. Rodsamran and R. Sothornvit
carboxymethyl cellulose-blended films (Rodsamran & Sothornvit, 2017)
and cassava starch films (Dos Santos Caetano et al., 2018). This was
because the hydrophilic nature of glycerol interacted with water or
biopolymer molecules forming hydrogen bonding. However, the co-
conut water (CW) concentration had no notable impact on the moisture
content of the lime peel pectin films (CW10 and CW20), probably due
to the relatively low amount of CW incorporated. No significant
changes in the moisture content of films were detected due to the ad-
dition of lime peel extract, except for G20E and CW20E. However, the
films containing lime peel extract had greater water solubility than the
films without lime peel extract. The high values of water solubility of
the films incorporated with lime peel extract might have been due to
the hydrophilic groups of phenolic compounds in the natural extract
that could easily interact with water molecules and thus improved the
hydrophilicity of the films (Liu et al., 2016; Riaz et al., 2018). Similar
trends had also been observed in gelatin films added with longan seed
extract (Sai-Ut, Benjakul, & Rawdkuen, 2015) and chitosan films in-
corporated with apple peel polyphenol extract (Riaz et al., 2018).
3.2. Opacity
Lime peel pectin films without natural extract were more trans-
parent compared to the films containing lime peel extract (Table 2).
Generally, it is desirable to produce transparent food packaging films in
order to show food products inside the package, whereas there is an
increasing demand to obtain light barrier films avoiding deteriorative
reactions (Sogut & Seydim, 2018). The addition of lime peel extract into
pectin films aimed to produce transparent films with higher opacity
values. Thus, the obtained films should be more suitable for those
packaging applications requiring product visibility. The current results
were similar to the addition of mango peel extract to gelatin films
(Adilah, Jamilah, Noranizan, & Hanani, 2018) and when grape seed
extract was added to chitosan films (Sogut & Seydim, 2018). Generally,
the opaqueness of biopolymer films was attributed to the interaction of
pectin matrix and polyphenol extract which reduced the light intensity
passing through the films.
3.3. Water vapor permeability
The WVP of lime peel pectin films incorporated with different
concentrations of natural lime peel extract ranged from 1.63 × 10−10
to 2.55 × 10−10 g⋅m−1s−1Pa−1. The films with glycerol (G20 and G30)
had similar WVP values, while the WVP of films with CW (CW10 and
CW20) significantly increased with increasing CW concentration.
Interestingly, the addition of lime peel extract in the films with CW
(CW10E and CW20E) lowered the WVP compared with the films
without the extract (CW10 and CW20). This decrease in WVP could
have been related to the hydrogen and covalent interactions between
the pectin matrix, functional groups of CW and polyphenolic com-
pounds of the extract, and consequently limited the availability of hy-
drogen groups to form hydrophilic bonds with water resulting in the
improvement in the film water barrier (Ganiari, Choulitoudi, &
Oreopoulou, 2017; Li, Miao, Wu, Chen, & Zhang, 2014). Similarly, the
incorporation of green tea extract and grape seed extract in alginate
films improved the film water barriers (Fabra, Falćo, Randazzo,
Sánchez, & López-Rubio, 2018). However, the addition of rosemary
extract to cassava starch films also increased WVP values (Piñeros-
Hernandez, Medina-Jaramillo, López-Córdoba, & Goyanes, 2017).
Moreover, the addition of spent coffee grounds increased WVP of pectin
films (Mendes et al., 2019). As known, the film compositions (hydro-
philic or hydrophobic nature) and concentration might attribute the
different behavior of film WVP values due to the intermolecular inter-
action among film matrix and the extract including the tortuosity
pathway of water molecules.
4
Food Hydrocolloids 97 (2019) 105173
3.4. Mechanical properties
The decrease in tensile strength (TS) and elastic modulus of films
resulted from increased amounts of plasticizers (either glycerol or CW)
and the addition of lime peel extract (Table 2). In fact, plasticizer
molecules and hydroxyl groups of the phenolic extract also interrupted
the formation of the pectin matrix resulting in a reduction in the TS and
elastic modulus. Similar results were reported by Norajit, Kim, and Ryu
(2010) for alginate films incorporated with ginseng extract and by Riaz
et al. (2018) for chitosan-based films containing apple peel extract.
Pectin film with mango or guava purees was also greatly lower tensile
strength and modulus and higher elongation (Viana et al., 2018). This
might probably be the dilution of film matrix and the plasticization
effect of sugars and aids from fruit purees (Viana et al., 2018). Never-
theless, the pectin films incorporated with spent coffee grounds in-
creased film modulus and elongation (Mendes et al., 2019).
Increasing the amount of either glycerol or CW also increased film
elongation. Basically, plasticizers are able to insert between polymer
chain spaces reducing the intermolecular attraction between polymer
chains and increasing film flexibility (Ng & Tan, 2015). However, using
CW as a plasticizer resulted in greater elongation compared to using
glycerol. The incorporation of lime peel extract caused a significant
increase in the elongation of the films with both plasticizers, which
might be attributed to the binding of the phenolic compounds and the
polymer matrix, resulting in an increase in the free volume inside the
film structure (Rodsamran & Sothornvit, 2018). Thus, the film com-
positions, film structure and the intermolecular interactions are im-
portant factors affecting the mechanical properties of films.
3.5. Antioxidant properties
The total phenolic content (TPC) and the scavenging activities
against the DPPH and ABTS radicals of the lime peel pectin films after
being dissolved in water and ethanol are presented in Table 3. With the
water system, the films with CW had a higher TPC than films with
glycerol due to the natural phenolic compounds present in CW. In
contrast, when the films were dissolved in ethanol, no TPC was de-
tected. Lime peel extract itself has high TPC and antioxidant activity
(Rodsamran & Sothornvit, 2019a). Therefore, films incorporated with
lime peel extract had higher TPC values in either water or ethanol
systems. Piñeros-Hernandez et al. (2017) reported that the TPC of
cassava starch films increased after the addition of rosemary extract.
Moreover, the TPC in gelatin films increased with the addition of
mango peel extract (Adilah et al., 2018).
As expected, the antioxidant activities of the films using DPPH assay
and ABTS assay increased with the addition of lime peel extract in both
the water and ethanol systems. Interestingly, a very high TPC and an-
tioxidant activity were detected when the films were dissolved in the
water system, perhaps because the large released amount of TPC from
the lime peel extract into the film extract solution was related to the
water solubility of pectin, allowing for rapid water penetration into the
film matrix and diffusion of the active compounds into the film extract
solution. Our finding was in line with Piñeros-Hernandez et al. (2017),
who also reported that the TPC of cassava starch films containing ro-
semary extract migrated quickly to water, while a very low amount of
TPC was observed by using ethanol as a medium.
3.6. Performance of lime peel pectin films in the retardation of soybean oil
oxidation
Taking into account the properties of the lime peel pectin films, the
CW10 and CW10E films showed higher TPC content than the films
plasticized with glycerol. Moreover, the natural phenolic compounds of
CW itself showed higher antioxidant activity of the films and this might
help to retard soybean oil oxidation. Therefore, the CW10 and CW10E
films were selected to evaluate their efficacy in the retardation of
P. Rodsamran and R. Sothornvit Food Hydrocolloids 97 (2019) 105173

Table 3
Total phenolic content (TPC) and antioxidant activity of lime peel pectin films, evaluated using DPPH and ABTS assays.
Antioxidant property Film sample∗

G20 G30 CW10 CW20 G20E G30E CW10E CW20E

∗∗
Water system
TPC (mg GAE/g film) 8.50d ± 1.55 7.33d ± 0.62 17.30c ± 1.44 16.40c ± 0.33 80.81a ± 1.41 75.15b ± 2.67 81.01a ± 0.26 76.16b ± 2.27
Antioxidant capacity
ABTS assay (μM Trolox/g 19.50c ± 1.26 22.18c ± 0.06 48.51bc ± 5.66 100.15b ± 25.00 597.98a ± 21.21 532.04a ± 4.72 543.14a ± 50.62 546.82a ± 49.21
film)
DPPH assay (μM Trolox/ 9.26c ± 2.16 6.92c ± 1.82 5.51c ± 0.22 8.15c ± 0.93 51.98a ± 0.86 40.22ab ± 0.44 43.50 ab
± 12.86 38.70b ± 4.25
g film)
Ethanol system∗∗∗
TPC (mg GAE/g film) 0.00d ± 0.00 0.00d ± 0.00 0.00d ± 0.00 0.00d ± 0.00 11.83c ± 2.06 28.60a ± 1.03 9.52c ± 1.02 25.14b ± 2.26
Antioxidant capacity
ABTS assay (μM Trolox/g 3.71d ± 0.19 4.08d ± 0.88 2.27d ± 0.69 3.96d ± 0.86 12.68c ± 0.72 20.59a ± 0.11 10.26c ± 1.80 17.51b ± 1.92
film)
DPPH assay (μM Trolox/ 0.90e ± 0.01 0.98e ± 0.11 1.02e ± 0.05 1.10e ± 0.05 1.99c ± 0.11 3.05a ± 0.02 1.71d ± 0.20 2.76b ± 0.12
g film)

Values are expressed as mean of two replicates ± standard deviation.

Different lowercase superscript letters in the same row indicate significantly different values (p < 0.05).
∗
The abbreviations of film samples are following; G means glycerol, CW means coconut water, E means lime peel extract and the numbers following the letters mean
the amount used. For example, G20, CW10 and CW10E represent films plasticized with 20% glycerol, 10% coconut water and 10% coconut water incorporated with
lime peel extract, respectively.
∗∗
Films were dissolved in water to prepare film extract solution for testing.
∗∗∗
Films were dissolved in ethanol to prepare film extract solution for testing.

Fig. 1. Polypropylene film (PP-film) and the selected films (CW10-film and
CW10E-film) on top, with their film sachets filled up with soybean oil and
without packaging as control shown below. CW10 and CW10E represent films
plasticized with 10% coconut water and 10% coconut water incorporated with
lime peel extract, respectively.
Fig. 2. Thiobarbituric acid reactive substances (TBARs) of soybean oil packed
in polypropylene film (PP-film) and the selected films (CW10-film and CW10E-
film) sachets during 30 days of storage. Error bars show ± standard deviation.
CW10 and CW10E represent films plasticized with 10% coconut water and 10%
coconut water incorporated with lime peel extract, respectively.

soybean oil oxidation during storage at 27 ± 2 °C (50 ± 5% RH) for

30 days (Fig. 1). oxidation observed by a lower peroxide value of soybean oil during
Changes in TBARs were expressed as the amount of secondary storage at 50 °C for 30 days (Wu et al., 2019). This was due to the
products of lipid oxidation (malonaldehyde and other TBA-reacting higher oxygen barrier property and more dense structure of bioactive
components) (Salgado, López-Caballero, Gómez-Guillén, Mauri, & films (Wu et al., 2019). Viana et al. (2018) recommended that the
Montero, 2013). The malonaldehyde level in the oil sample (0 day) was pectin films were suitable for sachet application. Therefore, it would be
initially 1.73 ± 0.23 mg MDA/kg oil. TBARs values of the control and interesting to apply larger sheets of film packaging on other food pro-
sample packed with PP-film sachet increased sharply after 6 days of ducts to determine any impact on food quality during storage in a fu-
storage (Fig. 2). TBARs increased stepwise but not significantly in the ture study.
soybean oil packed in CW10-film and CW10E-film sachets throughout
the whole storage period. These results were related to the antioxidant
4. Conclusion
properties of these films (Table 3). The antioxidant compounds of the
CW10-film and CW10E-film sachets might have retarded lipid oxidation
Antioxidant films were prepared by integrating coconut water and
by delaying hydroperoxide (primary lipid oxidation products) de-
lime peel extract as natural antioxidants into lime peel pectin films to
gradation (Benjakul et al., 2005; Salgado et al., 2013). Similar results
serve as new bioactive edible films. Coconut water is also known to act
were observed in the inhibition of lipid oxidation in fish patties of
as a natural plasticizer in lime peel pectin film formation. Furthermore,
sunflower protein concentrate films incorporated with clove essential
the film water barrier was improved by the combination of coconut
oil (Salgado et al., 2013). Moreover, gelatin films incorporated with
water as a plasticizer and lime peel extract. As more lime peel extract
longan seed extract also inhibited lipid oxidation of soybean oil during
was added, the films increased both their total phenolic content and
storage at 28 °C for 30 days (Sai-Ut et al., 2015). In addition, pomelo
antioxidant activities. The bioactive films with and without lime peel
peel flour films incorporated with tea polyphenol showed antioxidant
extract were effective at delaying lipid oxidation of soybean oil during
activity and antimicrobial activity and the bioactive films reduced oil
storage for 30 days. This study indicated that lime peel pectin films

5
P. Rodsamran and R. Sothornvit
containing coconut water and lime peel extract showed potential as
packaging film materials to extend the shelf-life of food products and to
reduce the use of synthetic plastic films.
Conflicts of interest
We have no conflicts of interest to declare.
Acknowledgements
The authors gratefully acknowledge the financial support of the
Kasetsart University Research and Development Institute, Bangkok,
Thailand.
References
Abid, M., Cheikhrouhou, S., Renard, C. M. G. C., Bureau, S., Cuvelier, G., Attia, H., et al.
(2017). Characterization of pectins extracted from pomegranate peel and their gelling
properties. Food Chemistry, 215, 318–325.
Abid, M., Renard, C. M. G. C., Watrelot, A. A., Fendri, I., Attia, H., & Ayadi, M. A. (2016).
Yield and composition of pectin extracted from Tunisian pomegranate peel.
International Journal of Biological Macromolecules, 93, 186–194.
Adilah, A. N., Jamilah, B., Noranizan, M. A., & Hanani, Z. A. N. (2018). Utilization of
mango peel extracts on the biodegradable films for active packaging. Food Packaging
and Shelf Life, 16, 1–7.
American Society of Testing and Materials (ASTM) International (1997). Standard test
method for tensile properties of thin plastic sheeting (D882–97). West Conshohocken, PA,
USA: ASTM International.
Benjakul, S., Visessanguan, W., Phongkanpai, V., & Tanaka, M. (2005). Antioxidative
activity of caramelisation products and their preventive effect on lipid oxidation in
fish mince. Food Chemistry, 90, 231–239.
Bermúdez-Oria, A., Rodríguez-Gutiérrez, G., Vioque, B., Rubio-Senent, F., & Fernández-
Bolaños, J. (2017). Physical and functional properties of pectin-fish gelatin films
containing the olive phenols hydroxytyrosol and 3,4-dihydroxyphenylglycol.
Carbohydrate Polymers, 178, 368–377.
Bernhardt, D. C., Pérez, C. D., Fissore, E. N., De'Nobili, M. D., & Rojas, A. M. (2017).
Pectin-based composite film: Effect of corn husk fiber concentration their properties.
Carbohydrate Polymers, 164, 13–22.
Dos Santos Caetano, K., Lopes, N. A., Costa, T. H., Brandelli, A., Rodrigues, E., Flôres, S.
H., et al. (2018). Characterization of active biodegradable films based on cassava
starch and natural compounds. Food Packaging and Shelf Life, 16, 138–147.
Espitia, P. J. P., Du, W. X., de Jesús Avena-Bustillos, R., de Fátima Ferreira Soares, N., &
McHugh, T. H. (2014). Edible films from pectin: Physical-mechanical and anti-
microbial properties-A review. Food Hydrocolloids, 35, 287–296.
Fabra, M. J., Falćo, I., Randazzo, W., Sánchez, G., & López-Rubio, A. (2018). Antiviral and
antioxidant properties of active alginate edible films containing phenolic extracts.
Food Hydrocolloids, 81, 96–103.
Falguera, V., Quintero, J. P., Jiménez, A., Muñoz, J. A., & Ibarz, A. (2011). Edible films
and coatings: Structures, active functions and trends in their use. Trends in Food
Science & Technology, 22, 292–303.
Farris, S., Schaich, K. M., Liu, L., Cooke, P. H., Piergiovanni, L., & Yam, K. L. (2009).
Development of polyion-complex hydrogels as an alternative approach for the pro-
duction of bio-based polymers for food packaging: A review. Trends in Food Science &
Technology, 20, 316–332.
Fishman, M. L., Chau, H. K., Hoagland, P. D., & Hotchkiss, A. T. (2006). Microwave-
assisted extraction of lime pectin. Food Hydrocolloids, 20, 1170–1177.
Galanakis, C. M. (2013). Emerging technologies for the production of nutraceuticals from
agricultural by-products: A viewpoint of opportunities and challenges. Food and
Bioproducts Processing, 91, 575–579.
Ganiari, S., Choulitoudi, E., & Oreopoulou, V. (2017). Edible and active films and coatings
as carriers of natural antioxidants for lipid food. Trends in Food Science & Technology,
68, 70–82.
Food Hydrocolloids 97 (2019) 105173
Li, J. H., Miao, J., Wu, J. L., Chen, S. F., & Zhang, Q. Q. (2014). Preparation and char-
acterization of active gelatin-based films incorporated with natural antioxidants.
Food Hydrocolloids, 37, 166–173.
Liu, J., Meng, C. G., Yan, Y. H., Shan, Y. N., Kan, J., & Jin, C. H. (2016). Protocatechuic
acid grafted onto chitosan: Characterization and antioxidant activity. International
Journal of Biological Macromolecules, 89, 518–526.
McHugh, T. H., Avena-Bustillas, R., & Krochta, J. M. (1993). Hydrophilic edible films:
Modified procedure for water vapor permeability and explanation of thickness ef-
fects. Journal of Food Science, 58(4), 899–903.
Mendes, J. F., Martins, J. T., Manrich, A., Sena Neto, A. R., Pinheiro, A. C. M., Mattoso, L.
H. C., et al. (2019). Development and physical-chemical properties of pectin film
reinforced with spent coffee grounds by continuous casting. Carbohydrate Polymers,
210, 92–99.
Ng, S. F., & Tan, S. L. (2015). Development and in vitro assessment of alginate bilayer
films containing the olive compound hydroxytyrosol as an alternative for topical
chemotherapy. International Journal of Pharmaceutics, 495, 798–806.
Nisar, T., Wang, Z. C., Yang, X., Tian, Y., Iqbal, M., & Guo, Y. (2018). Characterization of
citrus pectin films integrated with clove bud essential oil: Physical, thermal, barrier,
antioxidant and antibacterial properties. International Journal of Biological
Macromolecules, 106, 670–680.
Norajit, K., Kim, M.,K., & Ryu, G. H. (2010). Comparative studies on the characterization
and antioxidant properties of biodegradable alginate films containing ginseng ex-
tract. Journal of Food Engineering, 98, 377–384.
Oliveira, T.Í. S., Zea-Redondo, L., Moates, G. K., Wellner, N., Cross, K., Waldron, K., et al.
(2016). Pomegranate peel pectin films as affected by montmorillonite. Food
Chemistry, 198, 107–112.
Piñeros-Hernandez, D., Medina-Jaramillo, C., López-Córdoba, A., & Goyanes, S. (2017).
Edible cassava starch films carrying rosemary antioxidant extracts for potential use as
active food packaging. Food Hydrocolloids, 63, 488–495.
Riaz, A., Lei, S., Akhtar, H. M. S., Wan, P., Chen, D., Jabbar, S., et al. (2018). Preparation
and characterization of chitosan-based antimicrobial active food packaging film in-
corporated with apple peel polyphenols. International Journal of Biological
Macromolecules, 114, 547–555.
Rodsamran, P., & Sothornvit, R. (2017). Rice stubble as a new biopolymer source to
produce carboxymethyl cellulose-blended films. Carbohydrate Polymers, 171, 94–101.
Rodsamran, P., & Sothornvit, R. (2018). Bioactive coconut protein concentrate films in-
corporated with antioxidant extract of mature coconut water. Food Hydrocolloids, 79,
243–252.
Rodsamran, P., & Sothornvit, R. (2019a). Extraction of phenolic compounds from lime
peel waste using ultrasonic-assisted and microwave-assisted extractions. Food
Bioscience, 28, 66–73.
Rodsamran, P., & Sothornvit, R. (2019b). Microwave heating extraction of pectin from
lime peel: Characterization and properties compared with the conventional heating
method. Food Chemistry, 278, 364–372.
Sai-Ut, S., Benjakul, S., & Rawdkuen, S. (2015). Retardation of lipid oxidation using ge-
latin film incorporated with longan seed extract compared with BHT. Journal of Food
Science & Technology, 52(9), 5842–5849.
Salgado, P. R., López-Caballero, M. E., Gómez-Guillén, M. C., Mauri, A. N., & Montero, M.
P. (2013). Sunflower protein films incorporated with clove essential oil have po-
tential application for the preservation of fish patties. Food Hydrocolloids, 33, 74–84.
Seixas, F. L., Fukuda, D. L., Turbiani, F. R. B., Garcia, P. S., de O. Petkowicz, C. L.,
Jagadevan, S., et al. (2014). Extraction of pectin from passion fruit peel (Passiflora
edulis f. flavicarpa) by microwave-induced heating. Food Hydrocolloids, 38, 186–192.
Sogut, E., & Seydim, A. C. (2018). Development of chitosan and polycaprolactone based
active bilayer films enhanced with nanocellulose and grape seed extract.
Carbohydrate Polymers, 195, 180–188.
Srivastava, A., Greenspan, P., Hartle, D. K., Hargrove, J. L., Amarowicz, R., & Pegg, R. B.
(2010). Antioxidant and anti-inflammatory activities of polyphenolics from
Southeastern U.S. range blackberry cultivars. Journal of Agricultural and Food
Chemistry, 58, 6102–6109.
Viana, R. M., Sa, N. M. S. M., Barros, M. O., Borges, M. D. F., & Azeredo, H. M. C. (2018).
Nanofibrillated bacterial cellulose and pectin films added with fruit purees.
Carbohydrate Polymers, 196, 27–32.
Wu, H., Lei, Y., Zhu, R., Zhao, M., Lu, J., Xiao, D., et al. (2019). Preparation and char-
acterization of bioactive edible packaging films based on pomelo peel flours in-
corporating tea polyphenol. Food Hydrocolloids, 90, 41–49.