Integrity of edible Nano-coatings and its effects on quality of strawberries

subjected to simulated in-transit vibrations

Rajiv Dhital1, Prabesh Joshi1, Nathalie Becerra2, Arosha Umagiliyage1, Tan Chai3, Punit
Kohli2, Ruplal Choudhary1*
1
Department of Plant, Soil and Agricultural Systems, Southern Illinois University, Carbondale.
2
Department of Chemistry and Biochemistry, Southern Illinois University, Carbondale.
3
Department of Mechanical Engineering, Southern Illinois University, Carbondale.

*Corresponding Author: 1205 Lincoln Drive Room 176, Carbondale IL 62901, USA. Email:
[email protected]; Phone: 618 453 6985

ABSTRACT

Strawberries are a popular fruit with a pleasing color and flavor. However, its delicate tissue and
high sugar content makes it highly perishable with visible mold. In this study, we have attempted
to test feasibility of a new edible coating for extending shelf life of ‗Chandler‘ strawberries
subjected to simulated vibrations of local transportation. Six types of coatings were compared
based on the quality of treated berries. Curcumin and limonene were used as natural
antimicrobials and coatings were prepared from their liposomes and were over-coated with
methyl cellulose. One set of each coating type were subjected to the simulated vibration of local
transportation. The vibrated samples had lower shelf life than non-vibrated samples, indicating a
robust coating which remains intact during road vibrations is required. Based on the number of
berries with visible mold, limonene liposomes showed significantly lower fungal growth
compared to the control on the 14th day of storage. Titratable acidity and total phenolic contents
were also found to be higher in limonene coated strawberries compared to other coatings. Further
study is suggested to test liposome coatings of limonene with different particle size to improve
integrity of the coatings when strawberries are subjected to local transportation.
Keywords: Strawberry shelf life; edible coating; simulated local transportation; liposome;
limonene; curcumin.

1. Introduction

© 2017. This manuscript version is made available under the Elsevier user license
http://www.elsevier.com/open-access/userlicense/1.0/
Strawberries (Fragaria x ananassa) are a high demand fruit because of their pleasant aroma,
brilliant color, and delicious taste. They are also a good source of natural antioxidants, vitamins,
minerals and a significant amount of anthocyanins, flavonoids and phenolics (Rice-Evans and
Miller 1996, Heinonen, Meyer et al. 1998). The berries are harvested at full maturity in order to
maintain sensory (visual appearance, firmness, color) and nutritional (phytonutrients, minerals
and vitamins) qualities (Hernandez-Munoz, Almenar et al. 2008) . One of the most important
quality indicators in strawberries is the sugar to acid ratio, which characterizes degree of
sweetness and depends on the maturity, cultivar and weather conditions of berries (Pineli,
Moretti et al. 2011, Hu et al. 2012). Sugar to acid ratio in matured strawberries varies with the
variety, usually within a range from 7:1 for fruits regarded as sweet and 6:1 for fruits regarded as
acidic in taste (Wozniak et al. 1996). Due to its high respiration rate, soft texture and sensitivity
to temperature and mechanical shocks and vibrations, strawberries have postharvest shelf life
shorter than 1 week under ideal conditions at 0 ºC . This results in high degree of perishability to
several pathogens, which would in turn cause changes in pH, titratable acidity, total soluble
solids (TSS), loss in color, firmness and weight resulting in spoilage and, shortening the shelf
life.

Several attempts have been made to increase the postharvest quality of fruits and vegetables. The
most common method for maintaining quality and preventing decay is the use of low
temperatures under refrigeration (Han and Nie 2004, Hernandez-Munoz, Almenar et al. 2006).
Similarly, use of low temperatures and modified atmospheric packaging (MAP) in combination
with increased concentration of carbon dioxide (Manning 1993) and paraffin-based active
coatings by the use of essential oils for paper packaging of fruits and vegetables (Rodriguez et
al., 2007) are also in practice. These strategies, however, are expensive, time consuming, and
cause change in visual appearance of fruits and develop off-flavor in fruits (Ke, Zhou et al.
1994). The need of alternative methods to minimize the risk of undesirable biological,
physiochemical, and physiological changes of fruits and vegetables is desirable (Holcroft and
Kader 1999).

2
There has been an increasing trend in use of natural bioactive edible coatings composed of
polysaccharides, proteins, lipids, resins or of various composites in post-harvest preservation of
fruits and vegetables (Valencia-Chamorro, Perez-Gago et al. 2010). Edible coatings such
carnauba wax, whey proteins, gluten, shellac coatings, mucilage starch have exhibited beneficial
roles in maintaining quality of fruits and vegetables (Raghav et al., 2016). Apart from protection
of products from mechanical and microbiological damage to the fruits, these compounds have
shown to preserve post-harvest quality of fruits by preventing the loss of volatile compounds
(Perez‐ Gago, Rojas et al. 2002). These coatings are developed from natural sources and are
easily biodegradable, which meets the demand of consumers to have a safer food product
(Pavlath and Orts 2009). These coatings are of interest as coating materials due to their low-cost,
biodegradability, and solubility in water (Debeaufort, Quezada-Gallo et al. 1998).

Researchers have demonstrated that the use of plant based essential oils and phenolic compounds
as coating materials can be done in order to increase the shelf life, prevent microbial growth, and
to prevent nutrients loss from foods (Salmieri and Lacroix 2006). These compounds have shown
strong antimicrobial and antifungal properties, which makes them a natural alternative for the
prevention of pathogenic and spoilage organisms that may occur in foods (Lacroix 2007).

Limonene ((R)-(+)-para-Mentha-1,8-diene) is an essential oil extracted from lemon peels and

other citrus fruits (Moufida and Marzouk 2003). It is commonly used as a food additive or
flavoring agent and has a Generally Recognized as Safe (GRAS) status by US Food and Drug
Administration (EPA 1994). Similarly, it has exhibited fungicidal activities against Botrytis and
Asperigillus niger, the most common spoilage causing molds for fruit (Sharma and Tripathi
2008). A natural phenylpropanoid dimer Curcumin (1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-
heptadiene-3,5-dione) is the principle curcuminoid of turmeric Curcuma longa L. which is used
as a spice and traditional medicine in various parts of South and East Asia (Dogra, Choudhary et
al. 2015). Several studies suggest curcumin to be an effective anti-proliferative, anti-oxidant and
anti-inflammatory agent (Maheshwari, Singh et al. 2006). Like limonene, curcumin is also
considered as ―Generally Recognized as Safe‖ for application in food and pharmaceutical
formulations by US Food and Drug Administration.

3
Although having many food preservation qualities, the use of essential oils in food preservation
is limited due to certain drawbacks. These include high costs and potential toxicity to the
consumers. An approach to address these demerits while maintaining the efficacy of essential
oils and decreasing their dose would be the incorporation of these chemicals in a formulation of
edible coatings (Perdones, Escriche et al. 2016). A study by Sanchez-Gonzalez et al. (2011), in
development of an antibacterial composite films of Hydroxypropylmethyl cellulose (HPMC) and
chitosan mixed with different essential oils (Lemon, tea tree and bergamot) showed that the
antibacterial activity of chitosan was enhanced when it was used with a mixture of the polymer
and essential oil. Similarly, chitosan films incorporated with essential oils inhibited growth of the
Gram negative and positive bacteria Escherichia coli, Listeria monocytogenes and
Staphylococcus aureus.

Application of nanomaterials have shown to have a potential impact in a wide range of industries
(Michael 2004, Wang, Liang et al. 2004). In food industries too, the application of nano-
technology to enhance the quality of fruits has been used extensively (Yang, Li et al. 2010).
Fruits and vegetables with nano-packing have shown better physiochemical, sensory
physiological and preservation properties compared with normal packaging (Huang and Hu
2006, Li and Wang 2006, Li, Li et al. 2009). Liposomes are amphiphilic vesicles containing
polar heads and hydrophobic carbon tails basically designed by dispersal of phospholipids in
water (Bangham 1961). They can transport hydrophilic components by encapsulation in aqueous
phase and hydrophobic components in stable state by inserting into hydrophobic domains
(Brandl 2001, Shin, Chung et al. 2013). Size of the liposomes can be controlled to the order of
nanometers, providing desirable properties to deliver essential chemicals of hydrophilic and
hydrophobic nature. Their application in food preservation includes the encapsulation of
nutrients, proteins, enzymes, antimicrobials and flavors and controlled release in the food
environment to maintain food quality and prevent microbial spoilage (Makwana, Choudhary et
al. 2015).

Mechanical injuries during transportations of fruits and vegetables between the chain of
harvesting and consumption are one of the major causes of decay of fruits and vegetables
(Barchi, Berardinelli et al. 2002). Strawberries are highly prone to in-transit vibration damage

4
causing skin abrasion and bruising. From these abrasions and bruises on the tissues of berries,
microbes are able to enter inside which in turn causes the degradation of berries and reduce the
shelf life (Fischer, Craig et al. 1992). Significant losses due to damage caused by in-transit
vibration has been recorded in strawberries (Pierson, Allen et al. 1982).

In this study, fresh ripened berries were picked up from the local farms in Southern Illinois. The
berries were stored in a cold room at 4°C before any treatment and processing was done. Since
the phenolic compounds, vitamin B and vitamin C are sensitive to the higher storage
temperatures; refrigerated temperature at 4 °C is considered as safe level for storage.

The berries were treated with various types of natural coatings and subjected to simulated shock
and vibration. The effect of different natural edible coating formulations on shelf life of
strawberries was compared. The effects of mechanical shocks and vibrations of simulated road
transport on the quality of berries were analyzed. Physiochemical and nutritional quality of
treated berries were evaluated by the measurement of visible mold growth, total soluble solids,
pH, titratable acidity, and total phenolic content at different time intervals.

2. Materials and Methods

Fresh ripened strawberries 'Chandler' were purchased from local farms located in southern
Illinois. The berries were inspected for bruises, visual fungal growth, and decay. Uniform sized
berries were selected and stored at 4°C prior to coating and mechanical vibration experiments.
Steps for preparation of coating materials are shown in a flowchart form (Figure 8).

2.1. Preparation of phytochemical solutions

Curcumin solution with a concentration of 50mM was used as a coating material. Powdered
curcumin was initially dissolved in 2-3 drops of ethanol. For example, for the preparation of 25
ml curcumin solution with concentration of 50mM, 0.00046gm curcumin powder was weighed
and dissolved in ethanol followed by addition of 25 ml nano pure water.

5
Similarly, for the preparation of 25ml limonene solution with the concentration of 50 mM
0.01703 ml D-limonene was poured in a volumetric flask and desired volume was made by
pouring 25ml Nano- pure water.

2.2. Preparation of Methyl cellulose solution

Methyl cellulose solution at a concentration of 1.5% (1.5gm in 100ml) was used as a coating
material. Initially, powdered Methyl cellulose was dissolved in warm water at a temperature
around 50ºC for the complete dissolution. Then, followed by addition of Nano-pure water to
make up the desired volume.

2.3. Preparation of Curcumin and D- Limonene Liposomes

Thin film dehydration method was used for the preparation of lipid film. Briefly, a mixture of
dimyristoylphosphatidylcholine (DMPC), Polydiacetylene (PDA; 10,12-Pentacosadiynoic acid)
and N-hydroxysuccinimide (NHS) was dissolved in 25 mL of dichloromethane in a round
bottom flask. The solution was then subjected to rotary evaporation for 1 hour to evaporate the
solvent and bilayer film formation. The resulting film was dried overnight by placing the flask on
a vacuum pump. The film was then hydrated by the addition of 50 mM concentration of
phytochemicals (Curcumin or d-limonene) prepared in Nano-pure water. The resulting solution
was then left for sonication for 20 minutes for the complete detachment of the film. Further, the
solution was placed in a probe sonicator (VCX 500, Vibra-cell, Newtown, CT) at 76 °C for 15
minutes to produce small vesicles with diameter less than 110 nm. The solution was then filtered
through 0.45 µm nylon fiber to remove the lipid aggregates. Thus obtained liposomes were
collected in a vial and covered with aluminum foil. Liposomes were polymerized by irradiation
with a UV lamp emitting at 254 nm for approximately 2-5 min using a Pen Ray (UVGL-58,
Minerallight, Upland, CA) UV source (4.5 mW/cm2) in air. The polymerized liposome solution
was transferred into a dialysis membrane and dialysis was done for 48 hours changing the water
every 4 hours. The dialyzed liposomes were stored at 4°C and stability was observed for 7 days.

6
The liposomes were characterized and the results were presented in Dogra et al. (2015). A
diagram representing the preparatory steps for the liposomes is given in Figure 9.

2.4. Application of coating materials

For each coating material types, six sterile clam shell boxes each containing 20 uniform sized
berries were selected. The non-coated sample was used as a control. Two sets of coatings were
prepared for the analysis of quality parameters. The first set consists of coatings in which
curcumin was used, namely; Curcumin only, Curcumin liposomes, Methyl cellulose, bilayer
coating of Curcumin followed by Methyl cellulose and non-coated as control. The second set
consists of coatings in which D-limonene was used, namely; D-limonene only, D-Limonene
liposomes, Methyl cellulose, bilayer coating of D-Limonene followed by Methyl cellulose and
non-coated as control.

Berries were dipped in the solutions of coating materials for 10 minutes, air-dried in a UV
sterilized cabinet drier for 1 hour at 20 ºC, transferred into clamp shell boxes, and stored at 4ºC.
For the coatings in which bilayer of phytochemicals and Methyl cellulose were used, the berries
were initially dipped in the phytochemicals for 10 minutes, dried, dipped in Methyl cellulose
solution and stored at 4ºC.

2.5. Vibration tests

For each coating material, three boxes of coated strawberries were subjected to simulated
vibration tests immediately after drying, while the other three boxes were not subjected to
vibration tests. The system used to simulate the vibrations during local transportations consists of
a vibration shaker (Modal Shop 2060E) with a test platform, a digital signal generator, and an
amplifier (Fig. 1). A sweep sine signal controlled by the signal generator and the amplifier was
fed to the shaker. Based on the frequency components of the measured vibrations in a truck when

7
transporting strawberries locally, each sinusoidal signal was set to sweep from 2 to 80 Hz at an
acceleration level of 0.4 g (1 g = 9.8 m/s2). A piezoelectric accelerometer placed on the test
platform and a National Instrument data acquisition system were used to monitor the vibration
levels during the test. Each box of strawberry sample was subjected to vibrations for 2 hours at
room temperature. The non-vibrated samples were also kept in the same conditions of
temperature for 2 hours. The vibrated and non-vibrated strawberries were again stored at 4ºC
after the vibration tests.
2.6. Evaluation of fungal decay on strawberries

The stored strawberries were visually inspected for fungal decay for 14 days at different time
intervals (0, 2, 5, 9 and 14 days) after coating and vibration tests were performed. Day 0
corresponded to the day of treatment and vibration. Fungal decay percentage on the berries was
calculated. Fungal decay percentage is defined as the percentage (%) of strawberries which
showed the visual presence of one or more colonies of molds on their surface during storage to
the total number of berries.

2.7. Total soluble solids (TSSs), Titratable acidity (TA) and pH determinations

The quality parameters TSS, TA and pH of strawberries were measured at different time
intervals (0, 2, 5, 9 and 14 days). One fruit from each triplicate were taken and wrapped in a
sterile cheesecloth and squeezed with hands. The TSSs of the resulting juice were measured at
20ºC by a Brix refractometer (r2 mini, Reichert Analytical Instruments, Depew, NY). Similarly,
pH of the juice was measured by a pH meter (Corning pH/ion analyzer 350). TA was determined
by titrating the diluted juice (5ml juice diluted in 95ml distilled water) up to pH 8.2 using 0.1N
NaOH.

2.8. Total phenolic compound analysis

Fruit samples at different days of storage (0, 2, 5, 9 and 14 days) after coating and vibration tests
were taken. Briefly, a 1.5 g strawberry sample grinded in a mortar and pestle was weighted and
extracted with 20ml mixture of acetone, water and acetic acid (70:29.5:0.5 v/v). The samples

8
were vortexed for 1 hour at room temperature for complete extraction, followed by
centrifugation at 1640 g for 15 minutes at 20ºC. The supernatant was then filtered and allowed to
stand at room temperature for evaporation of solvent. The residue was then dissolved in distilled
water to a volume of 20 ml.

Total phenolic content of the extracted juice were determined by the use of Folin-Ciocalteu
reagent as per the method of Slinkard and Singleton (1977). The standard calibration curve was
prepared by using Gallic acid as a standard. The result was expressed as milligrams per liter of
Gallic acid equivalents (GAE) per 100 gm fresh weight.

2.9. Statistical analysis

Significant differences among different coatings, vibration, and days of storage on different
quality parameters were analyzed with one-way ANOVAs. The coatings and vibration treatments
were compared on the basis of fungal decay percentage, pH, total polyphenol content, total
soluble solids content and titratable acidity. The level of significance for all the analysis was
chosen to be 0.05. The means for different treatments were compared with least significant
difference (LSD) test. All statistical analyses were performed using JMP software package for
windows (JMP®, Version 12.2. SAS Institute Inc., Cary, NC, 2016).

3. Result and Discussion

3.1. Fungal decay

Due to their high physiological activities, strawberries are highly perishable fruits with a short
shelf-life. From the observations made up to 14 days, fungal decay percentage was found to
increase with the number of days of storage in each types of coatings and control. The effect of
days of storage showed significant effect (p < 0.05) on fungal decay % of strawberries. A gray
mold, Botrytis cinerea, is the most predominant mold that causes decay of strawberries (Harvey
and Pentzer 1960). The organism primarily infects the flower causing rot or remains dormant.
The dormant mold shows its activity when the concentration of sugars increases and favorable

9
environmental conditions are available either before or after harvesting of berries (Ayala-Zavala,
Wang et al. 2004).
Among the two sets of coatings designed, in Set 1, i.e. set of curcumin treated strawberries,
effect of vibration on fungal decay % was also found to be significant (p = 0.048). The vibrated
curcumin treated strawberries were found to have significantly higher fungal decay (45.5 %)
compared to the non- vibrated strawberries (27.625 %). This effect was more pronounced om the
14th day of storage (Fig. 7). For all coating treatments, significant difference was not observed
between vibrated and non-vibrated until the 9th days of storage. However, from 9th days onwards
all the vibrated samples were found to have significantly higher mold growth. The difference in
fungal decay % between vibrated and non-vibrated can be attributed to the change in integrity of
coatings by the simulated vibration. Mechanical damage due to vibration contributes to a
significant loss in perishable fruits and vegetables. In a study by Fischer et al. (1992), major
damage of strawberries due to simulated in-transit vibration of 5.0 to 10Hz was reported.
Mechanical damages caused by vibrations can affect the integrity of plasma cells of skin and
chemical contents of fruits resulting in change in bloom and tissue softening of fruits (Zhou, Su
et al. 2007).
Among the berries coated with phytochemicals, curcumin coatings showed significantly lower
fungal decay percentage compared to control and limonene coatings up to 2nd day of storage
(Fig. 2a). However, there was no significant difference in the fungal decay % after the 2nd day of
storage. Curcumin coatings showed a lower fungal decay at the 14th day of storage (Fig. 2a),
which was found to be significant compared to control. Fungal decay % among the treatment
formulations containing MC had higher mean value (40.78%) as compared with control samples
(36.41%) (Fig. 2b) which is in agreement with the study conducted by Perdones et al. (2016).
The bilayer coating of phytochemicals and MC did not show any significant reductions in fungal
decay percentage compared to control (Fig. 2c).
Liposome coated samples exhibited an elongated preservation effects, supported by significantly
lower visual mold growth compared to control during the 14th day of storage onwards (Fig. 2d).
Curcumin liposome coated samples exhibited significant reduction in fungal decay in 2nd, 5th and
14th days of storage compared to control (Fig. 2d). Meanwhile, on the 14th day of storage,
limonene liposome coated samples were found to be significantly (p = 0.0247) effective in

10
controlling fungal decay compared to control (Fig. 2d). This indicates that, for further extension
of shelf life, it is suggested to study the effects of liposome coatings.

3.2. TSS
Total soluble solid was found to be significantly higher in the Set 2 limonene treated strawberries
as compared to the Set 1 curcumin treated strawberries (p = 0.0301). Days of storage also
showed significant effect (p = 0.0239) on the TSS on each set of coatings designed (Fig. 3). No
significant difference in TSS was observed between vibrated and non-vibrated strawberry
samples. It was observed that there was an increase in TSS with days of storage. The change in
TSS content might be due to the solubilization of the cell wall polyuronides and hemicelluloses
in mature strawberries (Hernandez-Munoz, Almenar et al. 2008). TSS values of strawberries
generally ranges from 7-12% depending upon the genotypic characteristic of the fruit (Galletta,
Maas et al. 1995). High TSS values are generally accompanied by good flavor of strawberries
(Kader 1991).

3.3. pH
The limonene coated sample set was found to have significantly higher pH than curcumin coated
samples (p < 0.0001) and the control (p = 0.0132) (Fig. 4). The days of storage was shown to
have significant effect (p < 0.05) on the pH among the curcumin treated strawberries whereas the
effect was not apparent among the limonene treated and control strawberries. The pH of
limonene coated berries, decreased up to 5 days of storage but there is no significant increase on
the 14th day. The obtained results are in agreement with similar research conducted by Montero
(1996) who observed initial decreases followed by increases of pH during storage of
strawberries. For both vibrated and non-vibrated curcumin coated and control berries, the pH
increased throughout storage. However, there was no difference in pH among vibrated and non-
vibrated samples.
A similar study conducted by Zheng et al. (2007) found an increase in pH during storage, which
can be related to effects of increased O2 on the respiration rate of fruits.
3.4. Titratable acidity (TA)
Days of storage was shown to have significant effect (p = 0.0284) on TA content among the set
of both vibrated and non-vibrated limonene coated strawberries. There was an increasing trend

11
observed up to the 9th day of storage, and a non-significant decrease in the value at 14 days of
storage. No pronounced effect was observed in TA among the set of both vibrated and non-
vibrated curcumin coated and control strawberries. Titratable acidity content was found to be
significantly higher (p < 0.0001) in limonene as compared to curcumin treated strawberries (Fig.
5). This observation indicates a decrease in the fruit respiration metabolism resulting an increase
in the TA (de Oliveira, Magnani et al. 2014). TA is defined as the percentage of citric acid per
strawberry wet weight. The differences in TA content observed between the curcumin coated and
uncoated samples compared with limonene coated samples can be attributed to the increased
water loss (Hernandez-Munoz, Almenar et al. 2008). Decreased TA content can be related to the
decline in the organoleptic quality of the strawberries (Yang, Li et al. 2010).

3.5. Total Phenolic Content (TPC)

Limonene coated strawberries were found to have significantly higher (p = 0.0023) TPC as
compared to curcumin coated and control samples (Fig 6). There was an increasing trend in the
TPC content of the limonene coated berries up to 14 days of storage. The change in the
concentrations of phenolic compounds in fruits during storage can be attributed to cell structure
breakdown (Macheix and Fleuriet 1990). Meanwhile, in curcumin coated and control samples,
there was no significant increase in the TPC content from 2nd day to 5th day of storage, however
significant increase was observed 5nd to 9th days of storage (Fig 6). The days of storage was
found to have a significant effect (p < 0.0001) on TPC content of curcumin treated strawberries
whereas the effect was not seen in limonene treated strawberries. These observations concurred
with the findings by Ayala-Zavala et al. (2004).

4. Conclusion
In summary, the results obtained in this paper indicate that storage time significantly affects the
quality parameters of coated and non-coated strawberries. Vibration was also found to have a
significant effect on fungal decay and total soluble solids among all the coating treatments.
Among different coating types, liposomes were found to be the most effective for the
preservation of strawberry quality and the limonene liposome was found to be effective in
controlling fungal decay on strawberries for a prolonged period of storage. Similarly, titratable

12
acidity and total phenolic contents were also found to be higher in limonene coated strawberries
compared to other coatings.

Acknowledgement
Partial funding for this project came from Research Grant No. US-4680-13C from BARD The
United States - Israel Binational Agricultural Research and Development Fund; USDA-NIFA
Food Safety Outreach grant; National Institutes of Health; and the Elevating Research grant from
SIU, Carbondale, IL. April Vigardt provided trucks to measure vibrations during local produce
transportation, and Derek J. Fisher provide critical review of the manuscript.

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Figure Caption:
Figure 1. Experimental setup showing clamshell containing strawberry placed on the vibration
test apparatus

Figure 2a: Fungal decay % varying with the days of storage among the phytochemical treatments
only. Error bars represent ± Standard Error

Figure 2b. Fungal decay % varying with the days of storage among the strawberries coated with
methyl cellulose (MC) compared to control. Error bars represent ± Standard Error

Figure 2c. Fungal decay % varying with the days of storage among the coating treatments mixed
with methyl cellulose (MC). Error bars represent ± Standard Error

Figure 2d. Fungal decay % varying with the days of storage among the liposome coating
treatments compared to control. Error bars represent ± Standard Error

Figure 3. Mean total soluble solids for two sets of coatings. Error bars represent ± Standard
Error

Figure 4: Mean pH for two set of coatings. Error bars represent ± Standard error

Figure 5. Mean titratable acidity for two sets of coatings. Error bars represent ± Standard Error

Figure 6. Mean total phenolic content for two set of coatings. Error bars represent ± Standard
Error

Figure 7. Fungal decay percentage among vibrated and non-vibrated samples. Error bars
represent ± Standard Error

Figure 8. Flowchart representing steps in preparation of coating materials

Figure 9. Pictorial representation of liposome preparation

17
Figure 1. Experimental setup showing clamshell containing strawberry placed on the vibration
test apparatus.

18
 90
80
Fungal decay % 70
60
50
40
30
20
10
0
2 5 9 14
Days of storage

Curcumin only Limonene only Control

Figure 2a. Fungal decay % varying with the days of storage among the phytochemical treatments
only. Error bars represent ± Standard Error

19
 100
90
80
70
Fungal decay %

60
50
40
30
20
10
0
2 5 9 14
Days of storage

Control MC Only

Figure 2b. Fungal decay % varying with the days of storage among the strawberries coated with
methyl cellulose (MC) compared to control. Error bars represent ± Standard Error

20
 120

100
Fungal decay %

80

60

40

20

0
2 5 9 14
Days of storage

Curcumin with MC Limonene with MC Control

Figure 2c. Fungal decay % varying with the days of storage among the coating treatments mixed
with methyl cellulose (MC). Error bars represent ± Standard Error

21
 90
80
70
Fungal decay %

60
50
40
30
20
10
0
2 5 9 14
Days of storage
Curcumin liposome Limonene liposome Control

Figure 2d. Fungal decay % varying with the days of storage among the liposome coating
treatments compared to control. Error bars represent ± Standard Error

22
 12
Total Soluble Solids TSS (°Brix)

10

0
2 5 9 14
Days of storage

Curcumin Limonene Control

Figure 3. Mean total soluble solids for two sets of coatings. Error bars represent ± Standard
Error

23
 8
7
6
5
pH

4
3
2
1
0
2 5 9 14
Days of storage

Curcumin Limonene Control

Figure 4. Mean pH for two set of coatings. Error bars represent ± Standard error

24
 1.2

Titratable Acidity (g citric acid/100ml)

1

0.8

0.6

0.4

0.2

0
2 5 9 14
Days of storage

Curcumin Limonene Control

Figure 5. Mean titratable acidity for two sets of coatings. Error bars represent ± Standard Error

25
 2000

Total Phenolic content (mg GAE per

1800
1600

100gm fresh weight)

1400
1200
1000
800
600
400
200
0
2 5 9 14
Days of storage

Curcumin Limonene Control

Figure 6. Mean total phenolic content for two set of coatings. Error bars represent ± Standard
Error

Fungal decay on 14th day of storage

100
90
80
Fungal decay %

70
60
50
40
30
20
10
0
Control Curcumin Limonene

Vibrated Non-vibrated

Figure 7. Fungal decay percentage among vibrated and non-vibrated samples. Error bars
represent ± Standard Error

26
 Preparation of coating
materials

Preparation of 50 mM Preparation 50 mM Preparation of 1.5%

curcumin solution limonene solution methyl cellulose
solution
Curcumin powder Stock solution Methyl cellulose
powder
Weighed Limonene solution
of 50 mM Weighed
Dissolved in 2-3 Added with
drops of Ethanol appropriate Placed in warm
volume of nano water (50°C) until
pure water completely
Desired volume
made by addition dissolved
of nano pure water Limonene solution
of desired Addition of nano
concentration and pure water to
volume make desired
volume

Figure 8. Flowchart representing steps in preparation of coating materials

27
Figure 9. Pictorial representation of liposome preparation

28