International Journal of Life Sciences Biotechnology and Pharma Research Vol. 4, No.

2, April 2015

Antibacterial Active Packaging Edible Film

Formulation with Addition Teak (Tectona
grandis) Leaf Extract
Joni Kusnadi and Ponco Budyanto
Agriculture Product Technology, Faculty of Agricultural Technology, Brawijaya University, Jl. Veteran, Malang,
Indonesia 65145
Email: jkusnadi@ub.ac.id; joni.kusnadi@gmail.com, poncobudyanto@yahoo.co

production of PT. Perhutani of 800 thousand m3 per year,

however the utilization of teak leaves is still limited [6].
Addition of teak leaf extract as antibacterial material to
edible made from carrageenan and starch should capable
of extending the shelf life of the edible film and minimize
bacterial contamination of pathogenic and spoilage in
foods such as S. aureus and E. coli and also increase the
utilization of teak leafs.
Starch is a natural polymer that can readily be formed
into films. It consists of (1-4) linked -D-glucopyranosyl
units. Natural starch, amylopectin and amylose, consist of
this two kinds of chain. The linear polymer, amylose,
makes up about 20 wt% of the granule, and the remain 80
wt% consist of branched polymer, amylopectin. Amylose
is crystalline which has an average molecular weight of
500,000, while amylopectin is highly branched and have
higher molecular-weight [7]. Despite their ease of
preparation, starch films have poor physical properties.
These can be improved by blending with either synthetic
polymers to produce biodegradable materials [8] or other
natural polymers in edible packaging.
Carrageenan is a water soluble linear biopolymer,
increasingly used as natural thickener, formulation
stabilizer, or gelling agent in applications ranging from
food industry (mainly dairy products) to pharmaceutics
[9]. The dairy sector accounts for a large part of the
carrageenan applications in food products, such as frozen
desserts, chocolate milk, cottage cheese and whipped
cream. In addition to this, carrageenans are used in
various non-dairy food products, such as instant products,
jellies, pet foods, sauces, and non-food products, such as
pharmaceutical formulations, cosmetics and oil well
drilling fluid [10], [11]. Except starch, carrageenan is,
with pectin, the main natural gelling polysaccharide
extracted from plants or seaweeds and used as high-value
functional ingredient in foods, cosmetics and
pharmaceuticals [12].
Carrageenan isomers which already known were -, -,
and -carrageenan (-kappa, -lambda and -iota), which
has differences in the number and position of the ester
sulfate groups belongs to the repeating galactose units. carrageenan has one negative charge per disaccharide
with a tendency to form a strong but rigid gel. carrageenan has two and -carrageenan has an average of
2.7 charges per disaccharide unit. The gelling power of -

AbstractThe objective of this research is to investigate the

effect of teak leaf extract as antibacterial to the quality and
antibacterial activity of active packaging edible film. This
research is arranged used Randomized Block Design
consists of 2 factor combinations. First factor is carrageenan
concentration consist of 3, 5 and 7% of tapiocas weight.
Second factor is teak leaf extracts concentrations consist of
10, 15 and 20% of edible films solutions volume and
repeated 3 times. The data were analyzed with ANOVA and
continued with BNT or DMRT =5%. The best treatment
was determined using Multiple Atribute Method. The result
showed that the addition of 7% carrageenan and 20% teak
leaf extract was the best treatment. The edible film
produced had 0,183 mm thickness, 11,635 gram/m2, 24hour
water vapor transmission, 54,334 % elongation, 1,034 N/cm2
tensile strength and 0,600 cm and 0,650 cm antibacterial
activity toward E. coli and S. aureus, respectively.
Index Termsantibacterial, caragenan, edible film, tectona
grandis extract

I.

INTRODUCTION

Seventy percent of food poisoning cases in Indonesia,

caused by the bacterium Staphylococcus aureus,
Eschericia coli and Coliform because of unhygiene
packaging. So it requires packaging of food products that
capable of avoiding the risk of product contamination by
pathogenic and spoilage bacteria. Todate some techniques
have been developed in order to produce active
packaging which is environmentally friendly and
naturally active. Packaging usually contain antibacterial,
absorbent material O2, CO2 absorbent or enhancer [1].
Widely used packaging which can be applied as an
active packaging is edible films. Edible film is made of a
thin layer of edible material, formed on top of the food
component as a barrier to mass transfer, as a carrier of
food and improves food handling [2]. The material for
edible film can use materials such as tapioca, carrageenan
[3], [4] and glycerol. Antibacteria are compounds that
inhibit the activity of bacteria patogen. Antibacteria have
been widely used as bioactive compounds for edible film.
One of Antibacterial compound is phenol. Phenol is one
of compound found in the leaves of teak [5]. Average
Manuscript received January 9, 2015; revised March 9, 2015.
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carrageenan makes it capable of forming good film [13],

[14]. The formation of edible coating with good film
forming and mechanical properties can be done by
blending starch with -carrageenan. Starch must be
gelatinized first before blending with carrageenan
The purpose of this research is to investigate the effect
of teak leaf extract as antibacterial to the quality and
antibacterial activity of active packaging edible film. So,
using this edible film which inhibit the growth and kill
pathogens and spoilage bacteria in food will extend the
shelf life and maintain the quality of foods.
II.

suspension together with tapioca starch edible film during

heating and gelatinization [18]. Edible film thickness is
due because of the higher concentration of carrageenan
used. The increase of carrageenan will increase in total
dissolved solids present in the film-forming solution, so
after the drying process will produce thicker films. In
general, the thickness effect on tensile strength,
elongation, and water vapor transmission rate [4].

METHODES

A. Materials and Test Isolates

Materials used for the extraction of teak leaf
antibacterial are fresh Netherlands teak leaf. Materials
used for testing the antibacterial property of the edible
film produced in this researhc were E. coli ATCC 25 922,
S. aureus ATCC 29 213, NA and NB which obtained
from the Laboratory of Food Microbiology Department
of Agricultural Technology UBs Faculty of Agricultural
Technology.

Figure 1. Average elongation of edible film formed from varies

carrageenan concentration with addition of varies teakleaf extract
concentration.

B.

Elongation
Elongation is the maximum elongation when the edible
film start to tear of when it streched [2]. The use of
carrageenan in the production of edible film resulted in
the ability to bind water into a well that produced
elongation edible film [21]. The results showed 53.67%59.33% elongation (Fig. 2). Elongation chategorized
good when elongation more than 50% and less when the
elongation less than 10% [21].

B. Antibacterial Extraction
In this study, the antibacterial compound was extracted
from fresh teak leaves using a modified method of
Effendi [15], solvent extraction using distilled water and
and 80 Watt microwave for 1 minute.
Edible film was made by mixing 5 grams tapioca, 1 ml
glycerol, carrageenan with various concentration and 100
ml of distilled water. All mixed and heated (temperature
65-70 C) until the dough gelatinized. After the mixture
is cooled to a temperature of 40 C, teak leaf extract was
added. The mixture then spread on a mold, and dried.
After the edible film dried, it can be taken from the mold.
C. Parameters Used
The parameters used in this research were the total
phenol [16], thickness, tensile strength, film elongation
[17], water vapor transmission [18], the antibacterial
activity [19], the best treatment [20].
D. Data Analysis
The data were analyzed using Analysis of Varian
(ANOVA) followed by Least Significant Difference
(LSD) using 5% confidence interval or DMRT (Duncan
Multiple Range Test) with a confidence interval of 5%.
The best treatment will be taken by the method of
multiple attributes [20].
III.

Figure 2. Average thickness of edible film produced from varies

carrageenan concentration with addition of varies teakleaf extract
concentration.

The highest elongation obtained from carrageenan

concentration of 3% and teak leaf extract by 10%. In
carrageenan concentration 5 and 7% decrease in
elongation. This is presumably due to the increase in the
percentage of solids to water volume [21]. More
carrageenan, carrageenan molecules produce a stronger
matrix which caused the non-elastic so that elongation
decreased [4]. The addition of carvacrol can lower
elongation of edible film [22]. The addition of
carrageenan affect the reduced elongation [4]. Elongation
of edible film is due the higher the concentration of
carrageenan used. The carrageenan molecules will form a
stronger film matrix so the films lost its elasticity or
easily broken (brittle), and consequently the percentage

RESULT AND DISCUSSION

A.

The Thickness of Edible Film

The lowest thickness produced by 3% carrageenan and
10% leaf extract and the highest produced by carrageenan
treatment of 7% and 20% leaf extract (Fig. 1). The use of
carrageenan increased soluble solids, causing higher
thickness [21]. More carrageenan added will increase the
edible polymer films, increasing the viscosity of the

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of extension decreases. Other research showed that the

increase of breaking strength of the film will be followed
distantly by decrease in the percentage of elongation.
Edible films with the smallest elongation usually have a
stronger film formed.

value of tensile strength [18]-[23]. The tensile strength of

the material arising from the reaction between the
polymer bonding between atoms or secondary polymer
chains against external forces is given [23]. An increase
of tensile strength has been also reported when guar gum
(galactomannan) was added to chitosan films, although
further increase of concentration of galactomannan in the
mixtures led to reduction of TS values [24]. The addition
of agar into cassava starch induced an improvement in
EB and TS of cassava starch-based films [25]. Different
polysaccharide ratios can be used to improve mechanical
properties depending on the degree of interaction between
the molecules of the components.

C.

Water Vapor Transmission

The more the concentration of carrageenan and teak
leaf extract the lower water vapor transmission (Fig. 3).
From the results obtained showed that water vapor
transmission ranges from 10.832 g/m2.24 hour - 27.683 g
/ m2.24 hours.
Glycerol is a substance which can improve the
properties of edible film as a barrier to water vapor [21].
The lowest water vapor transmission is obtained from the
treatment of carrageenan concentration of 7% and 20%
leaf extract. The combination of carrageenan molecules,
tapioca starch and glycerol which binds both to the edible
films caused low water vapor transmission [21]. The
water vapor transmission of edible films depends on
many factors, such as the integrity of the film, the
hydrophilicehydrophobic ratio, the ratio between
crystalline and amorphous zones, and the polymeric chain
mobility [22]. One of the major purposes of edible films
is to block moisture transfer between the food and the
surrounding atmosphere, therefore the WVP should be as
low as possible. Permeability is the contribution of
diffusivity and solubility of the permeant through the
solid matrix, being this parameters changed by the
structure of the matrix when using the same permeant

Figure 4. Average tensile strenght of edible film formed from varies

carrageenan concentration with addition of varies teakleaf extract
concentration.

E.

Antibacterial Activity of Edible Film

The addition of teak leaf extract showed antibacterial
activity on Escherichia coli ATCC 25922 and
Staphylococcus aureus ATCC29213 proven by the
formation of clear zone diameter 0.12cm-1.26cm and
0.14cm-1.37cm. other research showed that teak leaf
extract added on edible film caused inhibition of
Eschericia coli and Staphylococcus aureus; Psedomonas
aeruginosa, Proteus mirabilis, Salmonella typhimurium,
Klebsiella pneumoniae, Escherichia coli and Serratia
marcescens [27].
Teak leaf extract has antibacterial activity against
Escherichia coli [26]. Antibacterial contained by edible
films containing teak leaf extract more effect on
Staphylococcus aureus ATCC 29 213 compared against
E.coli ATCC 25922. This is caused by the structure of the
cell wall of Gram-positive bacteria such as
Staphylococcus aureus is simpler than the cells of Gramnegative bacteria, Escherichia coli, so cause antibacterial
easier entry into cells of gram-positive bacteria. Grampositive bacteria have a cell wall of different sensitivity to
antibiotics, physical and enzyme treatment compared
with gram-negative bacteria which Gram-positive
bacteria more sensitive than Gram-negative antibacterial
[28]. In order antibacterial effect on Gram-negative
bacteria, antibacterial must penetrate the outer membrane
of bacterial sheath advance. After that, the antibacterial
entry through the cell wall and reaches past the
periplasmic enzyme serine protease. The enzyme
responsible for the biosynthesis of the cell wall [29].
Carrageenan with the concentration of 7% and 20%
leaf extract resulted in the largest inhibition zone (Fig. 5).
The higher the concentration of leaf extract the higher
the antibacterial antibacterial activity, meaning that the

Figure 3. Average water vapor transmission rate of edible film

produced from varies carrageenan concentration with addition of varies
teakleaf extract concentration.

D. Tensile Strength
Mechanical strength and extensibility are generally
required for a film to resist external stress and maintain
its integrity when applied to food products.Tensile
strength is the maximum tensile stress is acceptable
edible film to break up [21]. The results of research show
the tensile strength of edible films ranged 0.434 N / cm21,434 N / cm2.
Lowest tensile strength obtained from carrageenan
concentration of 3% and 10% identity leaf extract and the
highest tensile strength obtained from carrageenan
concentration of 7% and 20% leaf extract (Fig. 4). In
general, the addition of carrageenan will add to the strong
bond so the gel formed becomes stronger and caused
higher tensile strength. Some research showed that the
higher the concentration of carrageenan will increase the

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bacteria will be killed faster when higher concentrations

of antibacterial given [26]. Phenol is a major component
in teak leaf extract as antibakteri. Fenol can be damaged
by oxidation. The efectiveness of oxidation depending on
the amount of O2 in the air and the material properties of
carrageenan [6] transmission of O2, as the small molecule
of carrageenan fill a void in polymer matrix so as to
minimize O2 that affected the oxidized phenols. The
hydrophobic nature of O2 and hydrophilic structure
carrageenan resulted in O2 difficult to penetrate the
polymer carrageenan [30], so phenol in the network are
better protected if the edible film [31].

oily foods [34], etc., but also manufacturing soft capsules

[39], [40], and especially non-gelatin capsules [41].
Indeed, this protective barrier can also be used in food
domain in order to prevent the transfer of moisture, gases,
flavours, or lipids and thus to maintain or improve food
quality and to increase food product shelf life [42].
Another promising emerging technology that has been
applied to various biopolymers, including, carrageenanbased coating, is their use as antimicrobial agent carriers
in active packaging systems [14]-[43].
F.

Selection of Best Treatment

The best treatment is obtained by using the method
according to the Multiple Attribute [44]. Based on test
results, it is obtained with the use of the best treatment
karagenan 7% concentration and teak leaf extract 20%
(K3E20). It can be shown from the value of each
parameter which can be seen Table I.
TABLE I.

Figure 5. Average inhibition zone diameter of edible film formed from

varies carrageenan concentration with addition of varies teakleaf extract
concentration.

Teak
bark
effective
against
Listeria
monocytogenes
and
methicillin
resistant
Staphylococcus aureus. The antibacterial compound
of teak leaf is 5-hydroxy-1, 4- naphthalenedione
(Juglone) [32]. This compound has been found to be
inhibitory to oral pathogens, notably Streptococcus
mutans, Streptococcus sanguis, Porphyromonas
gingivalis and Prevotella intermedia [33], [34]
Another study showed the synergistic in-vitro
antibacterial activity to formulate new cost effective
antimicrobial agent for multi-drug resistant
organisms, based on the synergistic activity of
Tetracycline with methanol extract of teak. Other
research shows maximum synergistic activity
against different bacteria both Gram-positive and
Gram-negative species. The higher synergistic rate
was achieved against Salmonella typhimurium
(MTCC 98), Klebsiella pneumonia (MTCC 432),
and lowest synergistic shows against Pichia pastoris
(MTCC 34), Escherichia coli (MTCC 729). No
synergistic activity was observed in Citrobacter
freondii (MTCC 1658) [26]. The antibacterial
activity was also tested for leaf, bark and wood
extracts of teak against Staphylococcus aureus
(ATCC 25923), Klebsiella pneumoniae (ATCC
700603), hospital strains of Salmonella paratyphi
and Proteus mirabilis [35].

Parameter

Perlakuan Terbaik

Ketebalan

0,180 (mm)

Transmisi Uap Air

11,635 (gram/m2.24jam)

Elongasi
Tensile Strenght

54,330 (%)
1,030 (N/cm2)

Aktivitas
Antibakteri
terhadap Eschericia coli
ATCC 25922

6,000 (mm)

Aktivitas
Antibakteri
terhadap
Staphylococcus
aureus ATCC 29213

6,500 (mm)

IV.

CONCLUSION

The addition of carrageenan has a significant influence

on the thickness, water vapor transmission and tensile
strength edible film. Addition of teak leaf extract has
significant impact on the thickness, water vapor
transmission of edible film.
The best treatment is obtained on addition of
carrageenan concentration as much as 7% and teak leaf
extract as much as 20% which results in a thickness of
0.183 mm, 11.635 grams of water vapor transmission per
m2.24 hour, 54.334% elongation, 1,034N/cm2 tensile
strength, 0,600 cm inhibition zone against E. coli
ATCC25 922 and 0.650 cm inhibition zone S. aureus
ATCC29213.
ACKNOWLEDGMENT
The author would like to thanks Dr. Estri Laras
Arumingtyas for useful discussions.

The use of carrageenan as edible films and coatings

already covers various fields of the food industry such as
application on fresh and frozen meat, poultry and fish to
prevent superficial dehydration [36], ham or sausagecasings [37], granulation-coated powders, dry solid foods,

2015 Int. J. Life Sci. Biotech. Pharm. Res.

THE BEST TREATMENT OF EDIBLE FILM WITH ADDITION OF

CARRAGEENAN AND TEAK LEAF EXTRAC

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International Journal of Life Sciences Biotechnology and Pharma Research Vol. 4, No. 2, April 2015

Joni Kusnadi was born in Tanjungkarang,

Lampung, Indonesia dated June 12, 1962. He
completed the S1 in the Department of
Agricultural Product Technology, Faculty of
Agricultural Technology, Gadjah Mada
University in 1986 and graduated S2 in the
Department of Biothecnology, Gadjah Mada
University in 1999. S3 education was
completed in 2008, on the program of
agricultural science, Post Graduate UB. In
1988 until now he is a lecturer of Department Completed the S1 in the
Department of Agricultural Product Technology, Faculty of Agricultural
technology, Brawijaya University, Malang, Indonesia in the field of
Food Microbiology study. In the year 2009 until now, he is also the
head of the post graduate study program of agricultural product
technology at Brawijaya Univegrsity, Malang, Indonesia.

2015 Int. J. Life Sci. Biotech. Pharm. Res.

84

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