coatings

Article
Chitosan-Based Antimicrobial Coating for Improving
Postharvest Shelf Life of Pineapple
Indra Bhusan Basumatary 1 , Avik Mukherjee 1 , Vimal Katiyar 2 , Santosh Kumar 1, * and Joydeep Dutta 3, *

1 Department of Food Engineering and Technology, Central Institute of Technology Kokrajhar,

Kokrajhar 783370, Assam, India; [email protected] (I.B.B.); [email protected] (A.M.)
2 Department of Chemical Engineering, Indian Institute of Technology Guwahati,
Guwahati 781039, Assam, India; [email protected]
3 Functional Materials, Department of Applied Physics, School of Engineering Sciences, KTH Royal Institute of
Technology, AlbaNova Universitets Centrum, Hannes Alfvéns väg 12, 11419 Stockholm, Sweden
* Correspondence: [email protected] (S.K.); [email protected] (J.D.)

Abstract: Rapid postharvest losses and quality deteriorations in pineapple are major challenges to
growers and handlers. Chitosan-based coatings on fruit surfaces have gained importance in recent
years to enhance postharvest shelf life of the fruits. In this study, aloe vera gel was added as a natural
antioxidant in chitosan-based composite coating containing ZnO nanoparticles. The developed
formulation was applied on the surface of freshly harvested pineapple fruits. ZnO nanoparticles
were used as an antimicrobial agent. Coated pineapple fruits were evaluated for weight loss, total
soluble solids, titratable acidity, decay index, maturity index, and sensory attributes, including visual
appearance, periodically at 5 day interval during storage. The results showed that the coating of the





 fruit reduced weight loss by about 5%, and also delayed ripening and oxidative decay compared to
the uncoated fruit. Thus, the developed coating formulation is a promising sustainable solution to
Citation: Basumatary, I.B.;
reduce postharvest losses and to extend shelf life of pineapples.
Mukherjee, A.; Katiyar, V.; Kumar, S.;
Dutta, J. Chitosan-Based
Antimicrobial Coating for Improving
Keywords: biopolymer; chitosan; aloe vera; ZnO NPs; nanocomposite; shelf life; antimicrobial
Postharvest Shelf Life of Pineapple. coating; edible film and coating
Coatings 2021, 11, 1366. https://
doi.org/10.3390/coatings11111366

Academic Editor: Alexandra 1. Introduction

Muñoz-Bonilla Pineapple (Ananus comosus) is one of the popular tropical fruits mostly exported
by Costa Rica, Philippines, Brazil, Thailand, India, Indonesia, Chile, Ivory Coast, and
Received: 9 October 2021
South Africa, and its total production in India reached about 1.7 million tons in the year
Accepted: 2 November 2021
2019 [1]. Pineapples have a unique flavor and taste and are rich in nutrients and an-
Published: 8 November 2021
tioxidants, which make them attractive to consumers due to their great health benefits.
Pineapples are often introduced in the diet as a treatment or to prevent constipation be-
Publisher’s Note: MDPI stays neutral
cause they increase bowel movement, improve intestinal health, and clean the kidneys [2,3].
with regard to jurisdictional claims in
Pineapple is a perishable fruit with short postharvest shelf life mainly due to a high rate of
published maps and institutional affil-
water loss, respiration, senescence, and ethylene production resulting in rapid ripening
iations.
and deterioration [4,5]. Mostly fruits are packed and transported to destinations as quickly
as possible, or stored under controlled cooled conditions. Various technologies such as
modified atmosphere packaging (MAP) [6], synthetic wax coating [7], and salicylic acid
treatment [8,9] are used to enhance the shelf life of pineapple fruits. MAP is an expensive
Copyright: © 2021 by the authors.
technique requiring high capital cost, whereas the others involving synthetic chemicals can
Licensee MDPI, Basel, Switzerland.
potentially be harmful to consumers and for the environment in the long run [10].
This article is an open access article
In this context, application of biopolymer-based coatings on fruits and vegetables
distributed under the terms and
is gaining popularity for its efficacy in prolonging postharvest shelf life. Such coat-
conditions of the Creative Commons
Attribution (CC BY) license (https://
ings on fruit surfaces can effectively reduce weight loss, respiration, rate of maturity,
creativecommons.org/licenses/by/
and ripening [11–14]. Additionally, biopolymer-based coatings are generally non-toxic,
4.0/). biodegradable, biocompatible, environment-friendly, often edible, and may act as a carrier

Coatings 2021, 11, 1366. https://doi.org/10.3390/coatings11111366 https://www.mdpi.com/journal/coatings

Coatings 2021, 11, 1366 2 of 12

for active substances including antimicrobial and antioxidant agents [14]. The coating
forms a thin layer on the fruit surface that acts as a protective barrier against gases and
moisture, leading to a reduced rate of respiration and transpiration [15,16]. Several biopoly-
mers such as chitosan, starch, alginate, carrageenan, soy protein, zein, and gelatin have
been used for development of coating formulations for shelf-life extension of fruits [17–21].
Among them, chitosan is one of the well-studied biopolymers due to its inherent an-
timicrobial activity, abundant availability, low cost of manufacturing, good film-forming
properties and environmentally friendly nature (Figure 1) [1,22–25]. Chitosan (CS) is a
soluble form of chitin, and it is commercially obtained through partial deacetylation of
chitin, which is extracted from crustacean and fish processing wastes [26]. Chitosan has
inherent antimicrobial and antioxidant properties [14,27], but it is essential to enhance these
functionalities in chitosan-based coatings to achieve improved results leading to higher
shelf life of coated fruits. Improvement of shelf life of fruits such as guava [28], green
tomato [29], longan fruit [30], plum [31], and strawberry [32] by applying chitosan-based
coatings, alone or incorporated with active agents, has been reported in the literature.

Figure 1. Chemical characteristics and functional properties of chitosan.

Aloe vera gel is a natural antimicrobial agent effective in hampering microbial mem-
brane transportation [33,34]. Additionally, aloe vera gel has been used as a natural an-
tioxidant for centuries, and it has been applied as an edible coating on fruits to pre-
vent loss of moisture, to control respiratory rate and maturation, and to delay oxida-
tive browning [34,35]. Zinc oxide nanoparticles (ZnO NPs) are generally recognized as
safe (GRAS) substances and have been well-studied as antimicrobial nanofillers that
have tremendous potential as active agents in packaging films and coatings for food
preservation [19]. Nanocomposite zinc-oxide-chitosan formulations have been reported to
augment antimicrobial properties of packaging films with 63% reduction in bacterial counts
in okra (Abelmoschus esculentus) after 12 days storage in the ambient atmosphere [36].
Thus, in this study, to prolong postharvest shelf life, chitosan-based coating to pre-
vent moisture loss and firmness and control respiratory rate and maturation develop-
ment was supplemented by aloe vera gel for delayed oxidative browning and zinc oxide
(ZnO) nanoparticles to reduce microorganism proliferation, thus delaying spoilage. This
study focuses on the development of chitosan-based composite coating formulations for
pineapple fruit and evaluates their effectiveness in enhancing postharvest shelf life of the
coated/treated fruit. The effects of the coating treatment on physicochemical and sensory
attributes and deterioration of pineapple fruits were studied periodically during 15 days
storage in ambient conditions and reported herein.
Coatings 2021, 11, 1366 3 of 12

2. Materials and Methods

2.1. Raw Materials and Chemicals
Chitosan powder (≥90% deacetylation, low molecular weight; 50–200 KDa) was pro-
cured from Research Lab Fine Chem Industries, Mumbai, India. Hydrochloric acid (HCl),
sodium hydroxide (NaOH), acetic acid (CH3 COOH), and zinc oxide (ZnO) nanopowder
(particle size < 100 nm) were supplied by Sigma Aldrich, Mumbai, India. Fresh aloe vera
leaves were collected and brought to the laboratory, peeled, and then pulp was collected in
a beaker. Pulp was ground with a spatula and the obtained gel was filtered using a muslin
cloth and the filtrate was collected in a clean glass container and was termed as aloe vera
gel (AVG). Healthy pineapple fruits without any physical injury and decay were collected
from a fruit grower in Siliguri, India (latitude 26.7271◦ N and longitude 88.3953◦ E).

2.2. Preparation of Coating Formulation

Coating formulations for postharvest treatment of pineapple fruits were prepared
from chitosan (CH), aloe vera gel (AVG), and zinc oxide nanoparticles (ZnO NPs). Initially,
700 mL of chitosan solution (1%, w/v) was mixed with 1% acetic acid solution and the
mixture was continuously stirred at 500 rpm for 18 h on a magnetic stirrer (REMI, Mumbai,
India). Equal volume of the 140 mL prepared chitosan solution was poured in five separate
beakers. Next, 25% and 50% (w/w) of AVG were added and mixed well by continuous
stirring on a magnetic stirrer at 1000 rpm in each of the 2 pairs of beakers. The remaining
140 mL of the prepared chitosan solution was stored for later use for the reference chitosan
coating. Coating formulations containing 25% and 50% AVG were further reinforced with
1 wt.% ZnO NPs, and again mixed thoroughly under continuous stirring on a magnetic
stirrer at 1000 rpm for 15 min. Thus, five different coating formulations, namely CH
only, CH/AVG-25%, CH/AVG-50%, CH/AVG-25%/ZnO-1%, and CH/AVG-50%/ZnO-1%
were prepared as summarized in Table 1.

Table 1. Composition of chitosan-based coating formulations containing aloe vera gel and ZnO nanoparticles.

Coating Formulation CH% (w/v) AVG% (w/w) ZnO NPs% (w/w)

CH 1 0 0
CH/AVG-25 1 25 0
CH/AVG-50 1 50 0
CH/AVG-25/ZnO-1 1 25 1
CH/AVG-50/ZnO-1 1 50 1

2.3. Application of the Prepared Coating Formulation on Pineapple Fruits

Sixty pieces of fresh pineapple fruits of similar size, color, and maturity were col-
lected, thoroughly washed with tap water, and then sanitized by dipping in aqueous
NaOCl solution (200 ppm) for 2 min. The fruits were then dried under an electric fan in
ambient conditions. The washed and dried pineapple fruits were randomly distributed
into 6 groups, each group having 10 fruits for coating/treatment with all the prepared
formulations. Each group of pineapple fruits was coated/sprayed with the five different
prepared coating formulations using a nozzle sprayer (Master Airbrush, San Diego, CA,
USA). The sixth group of fruits was not coated and was considered as a control sample.
The coated fruits were dried under an electric fan for 3 h in ambient conditions (25 ◦ C
and 65% RH) to evaporate the excess moisture. After drying, each group of the pineapple
fruits was placed in cardboard boxes and stored under ambient conditions for 15 days,
and periodic physicochemical, sensory, and shelf-life observations were undertaken at
every 5-day interval. The experiment was conducted in a completely randomized de-
sign in a 6 × 4 factorial scheme, considering 6 treatments; control (without coating), CH,
CH/AVG-25, CH/AVG-50, CH/AVG-25/ZnO-1, and CH/AVG-50/ZnO-1, and 4 evalu-
ation periods; 0, 5, 10, and 15 days. Fruits were examined by periodic measurement of
Coatings 2021, 11, 1366 4 of 12

weight loss, total soluble solids (TSS), titratable acidity (TA), color change, browning, and
decay index.

2.4. Physicochemical and Sensory Analysis of Coated/Treated Pineapple Fruits

2.4.1. Weight Loss
The weight loss of the treated fruits was determined gravimetrically. The pineapple
fruit samples were weighed before the treatment/coating and expressed as Wi , and weight
after storage of the fruit at each time interval was expressed as Wf . The % weight loss of
the treated fruits was measured at each 5 day interval and determined using Equation (1):

Weight loss (%) = (Wi − Wf )/Wi × 100 (1)

where ‘Wi ’ and ‘Wf ’ are the initial and the final weight of the fruit samples, respectively.

2.4.2. Total Soluble Solids (TSS) Content, Titratable Acidity (TA), and Maturity
Index (TSS/TA)
Total soluble solids (TSS) content was measured by a digital refractometer (Atago,
Tokyo, Japan) and was expressed as percentage soluble solids. Titratable acidity (TA)
was determined by the titration of pineapple juice with 0.1 N NaOH to a pH of 8.1, and
expressed as grams of citric acid equivalents per liter of juice. Maturity index was expressed
as the ratio of TSS/TA as described in the literature [37].

2.4.3. Determination of Decay Index (DI)

The degree of decay of treated fruits was expressed by decay index (DI). Fungal or
bacterial growth symptoms on the fruit surface were observed visually by using a four-
point scale, where 0 = healthy fruit, 1 = one very small lesion (beginning of infection),
2 = one lesion less than 10 mm in diameter, 3 = several lesions or 25% of fruit infected,
4 = 26%–50% of the treated fruit surface infected and quantified using Equation (2):

∑ (DI rating × no. of fruits at DI rating level)

DI = (2)
Total number of treted fruits

2.4.4. Evaluation of Sensory Attributes

Sensory characteristics of the coated/treated pineapple fruits were determined ac-
cording to the method reported by Chen et al., 2019, with slight modifications [38]. Ten
trained panelists (five men and five women: age between 20–35 years old) participated in
the evaluation of sensory attributes such as sweetness, taste, odor, visual appearance, and
overall quality of the treated and untreated pineapple fruits during ambient storage. All the
panelists were pretrained to be familiar with the different sensory attributes of pineapple.
A nine-point hedonic scale was used; 1 = extremely poor, 3 = poor, 5 = acceptable, 7 = good,
9 = excellent, and a score of more than 5 was considered acceptable for consumption.

2.4.5. Change in Visual Appearance of the Treated Pineapple

Visual observation of the treated and untreated pineapple fruits was conducted by
taking photographs of the whole and cut pineapple fruits periodically during storage
under controlled lighting conditions. The color change, fungal growth, lesions, progress
of ripening, and finally the deterioration in the fruits were monitored and recorded. The
ripening stages of coated and uncoated/control pineapple fruits were determined by visual
observation of fruit skin color. Visual observation of the treated and untreated pineapple
fruits was conducted according to the method given by George et al., 2017, with slight
modification based on the nine-point hedonic scale (9: liked very much; 5: not liked or
disliked; 1: disliked very much) which was structured to express the degree of acceptability
for appearance. A score of more than 5 was considered visually acceptable.
Coatings 2021, 11, 1366 5 of 12

2.5. Statistical Analysis

All data were analyzed statistically by one-way ANOVA (p ≤ 0.05). Mean comparison
was performed individually for each day using Tukey’s test at significance level 0.05. All the
analysis was performed in triplicate, i.e., each analysis such as weight loss was performed
three times using three different samples from the same fruit, and factorial design of the
experiment was adopted each time. The data were processed using OriginPro 9.0 s.

3. Results and Discussion

3.1. Weight Loss of Treated Pineapple Fruits
The weight loss in treated pineapple fruits during storage at ambient conditions is
summarized in Table S1. It is evident from the results that application of only chitosan
coating reduces the weight loss of pineapple fruits. However, addition of aloe vera gel
in chitosan coating further enhanced the reduction in weight loss, which was directly
proportional to the concentration of aloe vera gel added in the chitosan coating. When the
pineapple fruits were treated with the CH/AVG coating formulation reinforced with ZnO
NPs, it reduced the weight loss even further (Figure 2A). The uncoated pineapple and that
treated with only CH coating showed 16.60% and 14.40% weight loss, respectively, whereas
CH/AVG-50- and CH/AVG-50/ZnO-1-coated fruits showed weight loss of only 11.56%
and 11.40%, respectively, after 15 days of storage. Thus, the coating (CH/AVG-50/ZnO-1)
reduced weight loss of coated pineapple fruits by 5.2% compared to the uncoated fruits.
Similar reduction in weight loss was also reported in mango fruits upon treatment with
chitosan-based coatings incorporated with titanium dioxide (TiO2 ) nanoparticles [11].
Many authors also reported the reduction of weight loss in green tomato, carrot, mango,
and guava upon coating with chitosan formulation [11,29,39–41].

Figure 2. Effects of coating formulations on (A) weight loss, (B) TSS contents, (C) TA, and (D) maturity index (TSS/TA) of
the treated and untreated pineapple fruit stored at ambient condition for 15 days.
Coatings 2021, 11, 1366 6 of 12

3.2. Total Soluble Solids (TSS) of the Treated Pineapple

Total soluble solids (TSS) mainly include sucrose, fructose, glucose, and other dis-
solved substances such as mineral, protein, and organic acids which are present in fruit
juice. TSS contents of the treated and untreated pineapple fruits were measured period-
ically by Brix value determined via a refractometer (Table S1). It was observed that TSS
of untreated fruit increased throughout the storage period (from 11.5 to 14.4%), whereas
in the coated fruits, the TSS values increased during the first 10 days and after 15 days
of storage, the TSS values were almost the same as recorded at the onset prior to the
coatings (TSS = 11.5%). Pineapple fruits coated with CH/AVG-50/ZnO-1 formulation
reduced loss in TSS by almost 2.97% (from 14.47% to 11.5%), compared to the uncoated
samples (Figure 2B). The increase in TSS concentration was perhaps due to loss of moisture
during storage. The reduced TSS loss in coated fruits was due to the slower hydrolysis of
carbohydrates to sugars, indicating slowing down of the respiration rate and metabolic
activities [41]. Chitosan-based coatings on fruit surfaces act as a selective barrier prevent-
ing oxygen (O2 ) and carbon dioxide (CO2 ) permeability [42]. The obtained results are in
agreement with the findings of other similar studies reported in the literature [30,40].

3.3. Titratable Acidity (TA) and Maturity Index of the Treated Pineapple
Citric and malic acids are the major organic acids found in pineapple fruits responsible
for the acidity, flavor, and taste of the fruit. Titratable acidity (TA) of coated and uncoated
fruits decreased throughout the storage period, but the reduction in TA of the coated
pineapples was comparatively lower (Figure 2C). At the end of 15 days storage, less
reduction in TA value of the coated fruits was due to utilization of the acids present in the
fruits as substrate during reverse glycolysis/respiration (Table S1). These results confirm
that the chitosan composite coatings on pineapple fruits slow down the rate of respiration
because of antioxidant activity of aloe vera gel and barrier properties of the coating upon
treatment on the fruit surface. However, as expected, no significant effect on TA with ZnO
NPs reinforcement in the coating was observed. The obtained results are similar to what
has been reported for other fruits coated with chitosan-based composite coatings, such as
strawberry [12], guava [13,40], and longan fruit [30].
Maturity of the coated and uncoated pineapple fruits increased throughout the storage
period, indicating the ripening process (Figure 2D). However, the increase in maturity
index in the coated pineapples was almost 27% lower (from 24.52 to 18.00) compared to
uncoated fruits after 15 days storage (Table S1). Incorporation of aloe vera gel and ZnO NPs
in the chitosan coating had a positive impact on delaying maturity and rate of ripening,
thus enhancing postharvest shelf life of pineapple fruits. Similar trends were reported in
the case of strawberry treated with chitosan/lemon essential oil composite coating [37].

3.4. Decay Index

Decay index of the treated and untreated pineapple fruits is shown in Table 2. The
treated and untreated pineapple fruits gradually deteriorated during storage under ambient
conditions. Initially, during the first 5 days of storage, no significant decay was observed
in any fruit samples, whereas, after 10 days of storage, decay started in a few groups
of pineapple samples. A few millimeter lesions were observed on the control, CH- and
CH/AV-25/ZnO-1-coated pineapple fruits. The CH/AVG-50/ZnO-1-coated pineapple
fruits showed only 25% decay compared to the control, whereas there was no decay
observed in CH/AVG-50-coated fruit even after 15 days storage. The lower decay index
in the case of chitosan coating containing aloe vera gel and ZnO NPs might be due to
antifungal and antibacterial activity of aloe vera gel [43] and ZnO nanoparticle [44], which
improved with increase in their concentrations. Similar results were also reported in our
previous study, in which ZnO NPs were shown to improve antimicrobial properties of
chitosan and agar-based nanocomposite films [19,36]. The antifungal and antibacterial
activities of ZnO nanoparticles have been reported, and four possible mechanisms are
suggested; (i) production of reactive oxygen species (ROS), (ii) loss of cellular integrity
Coatings 2021, 11, 1366 7 of 12

due to interaction of ZnO NPs with the microbial cell wall, (iii) release of Zn2+ ions, and
(iv) entry of ZnO NPs into the microbial cell [45].

Table 2. Decay index (DI) of the treated and untreated pineapple fruits during ambient storage of
15 days.

Treatments Decay Index (DI)

– Day-0 Day-5 Day-10 Day-15
Control 0 aA 0 aA 0.50 ± 0.12 bB 1.33 ± 0.2 cC
CH only 0 aA 0 aA 0.25 ± 0.07 bAB 0.66 ± 0.11 bBC
CH/AVG-25 0 aA 0 aA 0 aA 1.00 ± 0.10 bC
CH/AVG-50 0 aA 0 aA 0 aA 0 aA
CH/AV-25%/ZnO-1 0 aA 0 aA 0.50 ± 0.15 bB 0.66 ± 0.04 bBC
CH/AVG-50%/ZnO-1 0 aA 0 aA 0 aA 0.33 ± 0.09 bB
The values are expressed as means ± SD of triplicate assays, and the lower case and upper case letters in
superscript with the values indicate they are significantly different by LSD at p < 0.05 between storage times in
columns and treatments in rows, respectively.

3.5. Sensory Qualities

The results of sensory analysis of nanocomposite-coated and uncoated pineapple fruits
stored in ambient conditions for 15 days are presented in Table 3. Sweetness of pineapple
fruits increased during the entire period of storage in all the coated and uncoated groups of
pineapples mainly due to maturation and weight loss of the fruits (Figure S1A). The taste of
pineapple fruit consistently deteriorated for uncoated samples, and the score decreased to
4.2 at the end of 15 days storage. However, the taste characteristics were found to improve
in the case of CH/AVG-50 (6.1), CH/AVG-25/ZnO-1 (6.2), and CH/AVG-50/ZnO-1 (6.2)
coated pineapples, compared to the score on 0-day (5.8) (Figure S1B). Similar trends were
also observed in the case of odor of the treated and untreated pineapple fruits (Figure S1C).
The odor scores for CH/AVG-50-, CH/AVG-25/ZnO-1-, and CH/AVG-50/ZnO-1-coated
samples after 15 days were 6.2, 6.2, and 6.4, respectively, whereas that for the uncoated
sample was only 3.7 (Table 3). The deterioration in odor or off flavor as suggested by
the panelists in the case of uncoated and a few other coated pineapple fruits might be
due to microbial spoilage, which did not occur in the case of the fruits coated with the
above-mentioned coatings. Visual appearance scores of the coated and uncoated pineapple
fruits decreased throughout the 15-day storage period in all samples, but the fruits coated
with CH/AVG-50, CH/AVG-25/ZnO-1, and CH/AVG-50/ZnO-1 showed acceptable ap-
pearance even after 15 days (Figure S1D). Overall acceptance scores for all samples were
maintained during the first 5 days of storage, and gradually decreased thereafter. How-
ever, the overall acceptance scores for CH/AV-50-, CH/AV-25/ZnO-1-, and CH/AV-50/
ZnO-1-coated pineapple fruits were 7.2, 7.3, and 7.5, respectively, which were maintained
and comparable with that of 0-day (7.8) (Table 3). In conclusion, the pineapple fruits treated
with CH/AV-50, CH/AV-25/ZnO-1, and CH/AV-50/ZnO-1 composite coatings showed
better maintenance of all the sensory attributes even after 15 days of storage, which is
supported by findings of similar studies [46,47].
Coatings 2021, 11, 1366 8 of 12

Table 3. Values for sensorial attributes of coated and uncoated pineapple fruits stored at ambient condition for 15 days.

Treatments
Storage Period
Parameter CH/AV-25/ CH/AV-50/
(Days) Control CH Only CH/AV-25 CH/AV-50
ZnO-1 ZnO-1
0 5.72 ± 0.17 aA 5.72 ± 0.17 aA 5.72 ± 0.17 aA 5.72 ± 0.17 aA 5.72 ± 0.17 aA 5.72 ± 0.17 aA
5 5.81 ± 0.25 aAB 6.07 ± 0.22 aA 6.52 ± 0.20 aB 6.07 ± 0.12 aA 5.93 ± 0.18 aA 6.01 ± 0.16 aA
Sweetness
10 6.51 ± 0.17 abB 6.20 ± 0.21 aA 7.32 ± 0.09 bC 6.40 ± 0.12 abA 6.63± 0.13 abB 7.24 ± 0.20 bB
15 6.54 ± 0.18 aB 7.11 ± 0.12 abB 7.51 ± 0.20 bC 7.53 ± 0.15 bB 6.87 ± 0.13 abB 7.16 ± 0.22 abB
0 5.80 ± 0.11 aA 5.80 ± 0.11 aA 5.80 ± 0.11 aA 5.80 ± 0.11 aA 5.80 ± 0.11 aA 5.80 ± 0.11 aA
5 5.72 ± 0.09 aA 6.57 ± 0.13 bB 6.13 ± 0.07 aA 5.74 ± 0.09 aA 5.86 ± 0.14 aA 5.83 ± 0.12 aA
Taste
10 5.13 ± 0.12 aB 5.67 ± 0.07 aA 7.01 ± 0.09 cB 6.21 ± 0.07 bA 6.59 ± 0.08 bcB 7.75 ± 0.04 dB
15 4.29 ± 0.12 aC 5.05 ± 0.09 bC 5.73 ± 0.12 cA 6.13 ± 0.13 cA 6.22 ± 0.14 cA 6.23 ± 0.06 cA
0 5.60 ± 0.16 aA 5.60 ± 0.16 aA 5.60 ± 0.16 aA 5.60 ± 0.16 aA 5.60 ± 0.16 aA 5.60 ± 0.16 aA
5 5.57 ± 0.09 aA 5.82 ± 0.11 aA 5.75 ± 0.18 aA 5.83 ± 0.14 aA 5.85 ± 0.16 aA 5.67 ± 0.14 aA
Odor
10 5.43 ± 0.18 aA 6.58 ± 0.13 bB 6.23 ± 0.13 abA 6.33 ± 0.11 bA 6.28 ± 0.10 abA 6.21 ± 0.13 abA
15 3.76 ± 0.16 aB 5.06 ± 0.09 bA 5.32 ± 0.12 bA 6.26 ± 0.11 cA 6.25± 0.14 cA 6.47 ± 0.08 cB
0 9.00 ± 0.08 aA 9.00 ± 0.08 aA 9.00 ± 0.08 aA 9.00 ± 0.08 aA 9.00 ± 0.08 aA 9.00 ± 0.08 aA
Visual 5 8.25 ± 0.05 aB 8.07 ± 0.07 aB 8.16 ± 0.05 aB 8.13 ± 0.04 aB 7.84 ± 0.06 aB 7.81 ± 0.08 bB
appearance 10 6.51 ± 0.09 aC 7.21 ± 0.14 bC 7.63 ± 0.09 bcC 7.84 ± 0.04 cC 7.66 ± 0.05 bcB 7.8 ± 0.04 cB
15 4.24 ± 0.10 aD 5.16 ± 0.12 bD 6.20 ± 0.15 cD 7.24 ± 0.06 dD 7.63 ± 0.10 dB 7.55 ± 0.07 dB
0 7.80 ± 0.10 aA 7.80 ± 0.10 aA 7.80 ± 0.10 aA 7.80 ± 0.10 aA 7.80 ± 0.10 aA 7.80 ± 0.10 aA
Overall 5 8.18 ± 0.14 aA 7.83 ± 0.21 aA 8.18 ± 0.11 aA 8.15 ± 0.12 aA 7.90 ± 0.14 aA 7.95 ± 0.05 aA
acceptance 10 5.72 ± 0.12 aB 6.23 ± 0.22 aB 6.47 ± 0.11 bB 6.83 ± 0.15 bcB 7.25 ± 0.13 cA 7.25 ± 0.12 cA
15 4.13 ± 0.11 aC 5.65 ± 0.06 bB 6.77 ± 0.12 cB 7.22 ± 0.08 cB 7.30 ± 0.11 cA 7.54 ± 0.18 cA
The values are expressed as means ± SD of triplicate assays, and the lower case and upper case letters in superscript with the values
indicate they are significantly different by LSD at p < 0.05 between treatments in columns and storage times in rows, respectively.

3.6. Visual Observation of the Treated Pineapple Fruits

Visual observation of the treated pineapple was carried out to monitor the color
change, fungal growth, lesions, progress of ripening, and finally the deterioration in
fruits. The color of the skin of pineapple fruit is an excellent indicator to monitor the
ripening of the fruits. It has been observed that the color of the treated and untreated
pineapple changes from green to yellow and then to brown or orange during storage
(Figure 3). The uncoated (control) pineapple showed rapid color changes and attained
brown color more rapidly than the coated pineapple fruits. In the case of CH/AVG-50-,
CH/AVG-25/ZnO-1-, and CH/AVG-50/ZnO-1-coated pineapples, the yellow color and
fresh appearance were maintained even after 15 days of storage (Figure 3). Similar trends
were also observed in the internal tissues when fruits were cut (Figure 4). It is evident
from the figure that damage occurs in 20%–30% of the internal tissues in the case of
uncoated, CH-, and CH/AVG-25-coated fruits, whereas such damages are not visible in the
case of CH/AVG-50-, CH/AVG-25/ZnO-1-, and CH/AVG-50/ZnO-1-coated pineapples
(Figure 4). This is perhaps due to protective effects of the nanocomposite coatings
because they consist of antimicrobial and antioxidant agents (ZnO NPs and aloe vera
gel). In addition, the coatings help to reduce water loss, respiration rate, gaseous exchange,
and metabolic activities, thus extending the postharvest shelf life of the coated fruits. The
improvements achieved in postharvest life of pineapple fruits in this study by applying
sustainable coatings developed from chitosan, aloe vera gel, and ZnO NPs are comparable
to those reported in the literature using a synthetic commercial wax (Sta-Fresh 2952) [7].
Coatings 2021, 11, 1366 9 of 12

Figure 3. Effect of chitosan-based coatings on color, ripening, and maturity of pineapple fruits during 15 days storage under
ambient condition.

Figure 4. Effect of chitosan-based coatings on internal tissue damage and deterioration of pineapple fruits during 15 days
storage under ambient condition.
Coatings 2021, 11, 1366 10 of 12

4. Conclusions
In this study, novel and smart nanocomposite coatings were developed from chitosan,
aloe vera, and ZnO nanoparticles for prolonging postharvest life of pineapple fruit. Aloe
vera and ZnO nanoparticles were incorporated in chitosan matrix as active agents to further
increase the antimicrobial and antioxidant activities of chitosan. The developed coating
reduced weight loss by ~5.2%, TSS losses by ~2.97%, and maturity by ~27% in the coated
pineapple during 15 days of storage in ambient condition. Addition of aloe vera and ZnO
NPs into the chitosan coating not only slows down the fruit ripening but also maintains
the sensory attributes of pineapple fruit at acceptable levels. Thus, the developed coating
formulations can be a sustainable and smart solution to maintain postharvest quality and
extend shelf life of pineapples.

Supplementary Materials: The following are available online at https://www.mdpi.com/article/

10.3390/coatings11111366/s1, Figure S1: Effects of coating formulations on sensory attribute scores;
(A) sweetness, (B) taste, (C) odor, and (D) visual appearance of the treated and untreated pineapple
fruit during 15 days storage at ambient condition, Table S1: Values of weight loss, total soluble solid
(TSS), titratable acidity (TA), and maturity index (TSS/TA) of the coated and uncoated pineapple
fruits during storage for 15 days at ambient condition.
Author Contributions: Conceptualization, I.B.B., A.M., V.K. and S.K.; study design, I.B.B., A.M., V.K.
and S.K.; formal analysis, I.B.B. and S.K.; investigation, I.B.B., A.M., V.K., J.D. and S.K.; writing—original
draft preparation, I.B.B., A.M. and S.K.; writing—review and editing, S.K., V.K. and J.D.; supervision,
S.K. and V.K.; funding acquisition, S.K. and V.K. All authors have read and agreed to the published
version of the manuscript.
Funding: This research was funded by Department of Biotechnology (DBT), Ministry of Science
and Technology, Govt. of India with No. BT/COE/34/SP28408/2018 for providing necessary
financial support.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Experimental data generated at Department of Food Engineering
and Technology, Central Institute of Technology Kokrajhar, Assam (India) are available from the
corresponding author on request.
Acknowledgments: S.K. and V.K. acknowledge ‘Sunrise Career Project (Ref: NECBH/2019-20/173)’
under North East Centre for Biological Sciences and Healthcare Engineering (NECBH) Twinning
Outreach Programme hosted by Indian Institute of Technology Guwahati (IITG), Guwahati, Assam
funded by Department of Biotechnology (DBT), Ministry of Science and Technology, Govt. of India
with number BT/COE/34/SP28408/2018 for providing necessary financial support.
Conflicts of Interest: The authors declare no conflict of interest.

References
1. Leading Countries in Pineapple Production Worldwide in 2019. Available online: https://www.statista.com/statistics/298517
/global-pineapple-production-by-leading-countries/ (accessed on 4 November 2021).
2. Farid Hossain, M. Nutritional value and medicinal benefits of pineapple. Int. J. Nutr. Food Sci. 2015, 4, 84–88. [CrossRef]
3. Mohd Ali, M.; Hashim, N.; Abd Aziz, S.; Lasekan, O. Pineapple (Ananas comosus): A comprehensive review of nutritional
values, volatile compounds, health benefits, and potential food products. Food Res. Int. 2020, 137, 109675. [CrossRef] [PubMed]
4. Paull, R.E.; Chen, N.J. Chapter 17.4—Tropical fruits: Pineapples. In Controlled and Modified Atmospheres for Fresh and Fresh-Cut
Produce; Gil, M.I., Beaudry, R., Eds.; Academic Press: Cambridge, MA, USA, 2020; pp. 381–388.
5. Montero-Calderón, M.; Rojas-Graü, M.A.; Martín-Belloso, O. Effect of packaging conditions on quality and shelf-life of fresh-cut
pineapple (Ananas comosus). Postharvest Biol. Technol. 2008, 50, 182–189. [CrossRef]
6. Budu, A.S.; Joyce, D.C. Effect of modified atmosphere packaging on the quality of minimally processed pineapple cv. ‘Smooth
Cayenne’ fruit. J. Hortic. Sci. Biotechnol. 2015, 80, 193–198. [CrossRef]
7. Li, X.; Zhu, X.; Wang, H.; Lin, X.; Lin, H.; Chen, W. Postharvest application of wax controls pineapple fruit ripening and improves
fruit quality. Postharvest Biol. Technol. 2018, 136, 99–110. [CrossRef]
8. Lu, X.; Sun, D.; Li, Y.; Shi, W.; Sun, G. Pre- and post-harvest salicylic acid treatments alleviate internal browning and maintain
quality of winter pineapple fruit. Sci. Hortic. 2011, 130, 97–101. [CrossRef]
Coatings 2021, 11, 1366 11 of 12

9. Sayyari, M.; Babalar, M.; Kalantari, S.; Serrano, M.; Valero, D. Effect of salicylic acid treatment on reducing chilling injury in
stored pomegranates. Postharvest Biol. Technol. 2009, 53, 152–154. [CrossRef]
10. Youryon, P.; Supapvanich, S.; Kongtrakool, P.; Wongs-Aree, C. Calcium chloride and calcium gluconate peduncle infiltrations
alleviate the internal browning of Queen pineapple in refrigerated storage. Hortic. Environ. Biotechnol. 2018, 59, 205–213.
[CrossRef]
11. Xing, Y.; Yang, H.; Guo, X.; Bi, X.; Liu, X.; Xu, Q.; Wang, Q.; Li, W.; Li, X.; Shui, Y.; et al. Effect of chitosan/Nano-TiO2 composite
coatings on the postharvest quality and physicochemical characteristics of mango fruits. Sci. Hortic. 2020, 263, 109135. [CrossRef]
12. Riaz, A.; Aadil, R.M.; Amoussa, A.M.O.; Bashari, M.; Abid, M.; Hashim, M.M. Application of chitosan-based apple peel
polyphenols edible coating on the preservation of strawberry (Fragaria ananassa cv Hongyan) fruit. J. Food Process. Preserv. 2020,
45, e15018. [CrossRef]
13. Hong, K.; Xie, J.; Zhang, L.; Sun, D.; Gong, D. Effects of chitosan coating on postharvest life and quality of guava (Psidium
guajava L.) fruit during cold storage. Sci. Hortic. 2012, 144, 172–178. [CrossRef]
14. Basumatary, I.B.; Mukherjee, A.; Katiyar, V.; Kumar, S. Biopolymer-based nanocomposite films and coatings: Recent advances in
shelf-life improvement of fruits and vegetables. Crit. Rev. Food Sci. Nutr. 2020, 1–24. [CrossRef] [PubMed]
15. Kumar, S.; Ye, F.; Dobretsov, S.; Dutta, J. Chitosan nanocomposite coatings for food, paints, and water treatment applications.
Appl. Sci. 2019, 9, 2409. [CrossRef]
16. Salehi, F. Edible coating of fruits and vegetables using natural gums: A review. Int. J. Fruit Sci. 2020, 20, S570–S589. [CrossRef]
17. Kumar, S.; Mudai, A.; Roy, B.; Basumatary, I.B.; Mukherjee, A.; Dutta, J. Biodegradable hybrid nanocomposite of chitosan/gelatin
and green synthesized zinc oxide nanoparticles for food packaging. Foods 2020, 9, 1143. [CrossRef] [PubMed]
18. Kumar, S.; Mitra, A.; Halder, D. Centella asiatica leaf mediated synthesis of silver nanocolloid and its application as filler in
gelatin based antimicrobial nanocomposite film. LWT 2017, 75, 293–300. [CrossRef]
19. Kumar, S.; Boro, J.C.; Ray, D.; Mukherjee, A.; Dutta, J. Bionanocomposite films of agar incorporated with ZnO nanoparticles as an
active packaging material for shelf life extension of green grape. Heliyon 2019, 5, e01867. [CrossRef] [PubMed]
20. Díaz-Mula, H.M.; Serrano, M.; Valero, D. Alginate coatings preserve fruit quality and bioactive compounds during storage of
sweet cherry fruit. Food Bioprocess Technol. 2012, 5, 2990–2997. [CrossRef]
21. Moreno, M.A.; Orqueda, M.E.; Gómez-Mascaraque, L.G.; Isla, M.I.; López-Rubio, A. Crosslinked electrospun zein-based food
packaging coatings containing bioactive chilto fruit extracts. Food Hydrocoll. 2019, 95, 496–505. [CrossRef]
22. Al-Naamani, L.; Dobretsov, S.; Dutta, J. Chitosan-zinc oxide nanoparticle composite coating for active food packaging applications.
Innov. Food Sci. Emerg. Technol. 2016, 38, 231–237. [CrossRef]
23. Kumar, S.; Mukherjee, A.; Dutta, J. Chitosan based nanocomposite films and coatings: Emerging antimicrobial food packaging
alternatives. Trends Food Sci. Technol. 2020, 97, 196–209. [CrossRef]
24. Kumar, S.; Shukla, A.; Baul, P.P.; Mitra, A.; Halder, D. Biodegradable hybrid nanocomposites of chitosan/gelatin and silver
nanoparticles for active food packaging applications. Food Packag. Shelf Life 2018, 16, 178–184. [CrossRef]
25. Kerch, G. Chitosan films and coatings prevent losses of fresh fruit nutritional quality: A review. Trends Food Sci. Technol. 2015, 46,
159–166. [CrossRef]
26. Boudouaia, N.; Bengharez, Z.; Jellali, S. Preparation and characterization of chitosan extracted from shrimp shells waste and
chitosan film: Application for eriochrome black T removal from aqueous solutions. Appl. Water Sci. 2019, 9, 91. [CrossRef]
27. Kumar, S.; Ye, F.; Mazinani, B.; Dobretsov, S.; Dutta, J. Chitosan nanocomposite coatings containing chemically resistant ZnO–SnOx
core–shell nanoparticles for photocatalytic antifouling. Int. J. Mol. Sci. 2021, 22, 4513. [CrossRef]
28. Nair, M.S.; Saxena, A.; Kaur, C. Effect of chitosan and alginate based coatings enriched with pomegranate peel extract to extend
the postharvest quality of guava (Psidium guajava L.). Food Chem. 2018, 240, 245–252. [CrossRef]
29. Zhu, Y.; Li, D.; Belwal, T.; Li, L.; Chen, H.; Xu, T.; Luo, Z. Effect of nano-SiOx/chitosan complex coating on the physicochemical
characteristics and preservation performance of green tomato. Molecules 2019, 24, 4552. [CrossRef]
30. Shi, S.; Wang, W.; Liu, L.; Wu, S.; Wei, Y.; Li, W. Effect of chitosan/nano-silica coating on the physicochemical characteristics of
longan fruit under ambient temperature. J. Food Eng. 2013, 118, 125–131. [CrossRef]
31. Kumar, P.; Sethi, S.; Sharma, R.R.; Srivastav, M.; Varghese, E. Effect of chitosan coating on postharvest life and quality of plum
during storage at low temperature. Sci. Hortic. 2017, 226, 104–109. [CrossRef]
32. Pavinatto, A.; de Almeida Mattos, A.V.; Malpass, A.C.G.; Okura, M.H.; Balogh, D.T.; Sanfelice, R.C. Coating with chitosan-based
edible films for mechanical/biological protection of strawberries. Int. J. Biol. Macromol. 2020, 151, 1004–1011. [CrossRef]
33. Goudarzi, M.; Fazeli, M.; Azad, M.; Seyedjavadi, S.S.; Mousavi, R. Aloe vera Gel: Effective therapeutic agent against multidrug-
resistant pseudomonas aeruginosa isolates recovered from burn wound infections. Chemother. Res. Pract. 2015, 2015, 639806.
[CrossRef]
34. Hassanpour, H. Effect of Aloe vera gel coating on antioxidant capacity, antioxidant enzyme activities and decay in raspberry fruit.
LWT—Food Sci. Technol. 2015, 60, 495–501. [CrossRef]
35. Misir, J.; Brishti, F.H.; Hoque, M.M. Aloe vera gel as a novel edible coating for fresh fruits: A review. Am. J. Food Sci. Technol. 2014,
2, 93–97. [CrossRef]
36. Al-Naamani, L.; Dutta, J.; Dobretsov, S. Nanocomposite zinc oxide-chitosan coatings on polyethylene films for extending storage
life of okra (abelmoschus esculentus). Nanomaterials 2018, 8, 479. [CrossRef] [PubMed]
Coatings 2021, 11, 1366 12 of 12

37. Perdones, A.; Sánchez-González, L.; Chiralt, A.; Vargas, M. Effect of chitosan–lemon essential oil coatings on storage-keeping
quality of strawberry. Postharvest Biol. Technol. 2012, 70, 32–41. [CrossRef]
38. Chen, H.; Sun, Z.; Yang, H. Effect of carnauba wax-based coating containing glycerol monolaurate on the quality maintenance
and shelf-life of Indian jujube (Zizyphus mauritiana Lamk.) fruit during storage. Sci. Hortic. 2019, 244, 157–164. [CrossRef]
39. Sanuja, S.; Agalya, A.; Umapathy, M.J. Synthesis and characterization of zinc oxide-neem oil-chitosan bionanocomposite for food
packaging application. Int. J. Biol. Macromol. 2015, 74, 76–84. [CrossRef]
40. Batista Silva, W.; Cosme Silva, G.M.; Santana, D.B.; Salvador, A.R.; Medeiros, D.B.; Belghith, I.; da Silva, N.M.; Cordeiro, M.H.M.;
Misobutsi, G.P. Chitosan delays ripening and ROS production in guava (Psidium guajava L.) fruit. Food Chem. 2018, 242, 232–238.
[CrossRef]
41. Rohani, M.; Zaipun, M.; Norhayati, M. Effect of modified atmosphere on the storage life and quality of Eksotika papaya. J. Trop.
Agric. Food Sci. 1997, 25, 103–114.
42. Elsabee, M.Z. Chitosan-Based Edible Films. In Polysaccharides: Bioactivity and Biotechnology; Ramawat, K.G., Mérillon, J.-M., Eds.;
Springer International Publishing: Cham, Switerland, 2015; pp. 829–870.
43. Abakar, H.O.M.; Bakhiet, S.E.; Abadi, R.S.M. Antimicrobial activity and minimum inhibitory concentration of Aloe vera sap and
leaves using different extracts. J. Pharmacogn. Phytochem. 2017, 6, 298–303.
44. Kaushik, M.; Niranjan, R.; Thangam, R.; Madhan, B.; Pandiyarasan, V.; Ramachandran, C.; Oh, D.-H.; Venkatasubbu, G.D.
Investigations on the antimicrobial activity and wound healing potential of ZnO nanoparticles. Appl. Surf. Sci. 2019, 479,
1169–1177. [CrossRef]
45. Lallo da Silva, B.; Abucafy, M.P.; Berbel Manaia, E.; Oshiro Junior, J.A.; Chiari-Andreo, B.G.; Pietro, R.C.R.; Chiavacci, L.A.
Relationship between structure and antimicrobial activity of zinc oxide nanoparticles: An overview. Int. J. Nanomed. 2019, 14,
9395–9410. [CrossRef] [PubMed]
46. Guimarães, G.; Dantas, R.; Sousa, A.; Soares, L.; Raylson, D.; Rosana, S.; Lima, R.; Rejane, M.; Beaudry, R.; Silva, S. Impact of
cassava starch-alginate based coatings added with ascorbic acid and elicitor on quality and sensory attributes during pineapple
storage. Afr. J. Agric. Res. 2017, 12, 664–673. [CrossRef]
47. Mandal, D.; Lalremruata; Hazarika, T.; Nautiyal, B.P. Effect of post-harvest treatments on quality and shelf life of pineapple
(Ananas comosus [L.] Merr. 'Giant Kew') fruits at ambient storage condition. Int. J. Bio-Resour. Stress Manag. 2015, 6, 490–496.
[CrossRef]