American Journal of Food Science and Nutrition Research

2017; 4(6): 178-183

http://www.openscienceonline.com/journal/fsnr
ISSN: 2381-621X (Print); ISSN: 2381-6228 (Online)

The Utilization of Chitosan from Shrimp Shells as

Edible Coating for Wooden Fish (Keumamah)
Resmila Dewi, Endang Sutariningsih Soetarto
Faculty of Biology, University of Gadjah Mada, Yogyakarta, Indonesia

Email address
resmila_dewi@yahoo.com (R. Dewi)

To cite this article

Resmila Dewi, Endang Sutariningsih Soetarto. The Utilization of Chitosan from Shrimp Shells as Edible Coating for Wooden Fish
(Keumamah). American Journal of Food Science and Nutrition Research. Vol. 4, No. 6, 2017, pp. 178-183.

Received: April 11, 2017; Accepted: August 17, 2017; Published: September 25, 2017

Abstract
Wooden fish (keumamah) is a traditional food made from tuna (Euthynnus affinis) coated with wheat flour and be drained.
During the process of storage and marketing of wooden fish is often contaminated by certain types of mold, especially
Aspergillus sp. The presence of mold contaminants would reduce the wooden fish quality and dangerous for consumption. It is
therefore necessary natural ingredients that can act as a preservative, one of which is chitosan. The purpose of this study was to
analyze the long wooden fish optimum soaking in a solution of chitosan to produce wooden fish that have a longer shelf life
than fish-covered wood flour. The study begins by preparing chitosan from shrimp shells. Chitosan is extracted from shrimp
shells, with a solution of NaOH and HCl and dried at a temperature of 65°C. Wooden fish with chitosan coating is done by
soaking the fish in a timber with 1.5% chitosan larutam time variation of 0, 15, 30, 45, 60 and 75 minutes. The fish dried in the
sun to harden fish such as timber, stored at room temperature (± 30°C). Test storability of wooden fish is done until a certain
time limit (4 weeks), namely on days 0, 7, 14, 21, and 28, through organoleptic (color, texture, aroma and taste) and mold on
the fish population analysis of timber. The results showed that soaking the fish for 60-75 minutes in a solution of chitosan 1.5%
(w/v) to produce the most good organoleptic value, with no evidence of mold growth on the shelf life of 28 days so that the
wooden fish unfit for consumption.

Keywords
Chitosan, Wooden Fish, Edible Coating, Mold

been contaminated by pathogenic fungi or products containing

1. Introduction
Wooden fish (keumamah) is processed tuna (Euthynnus
affinis) were coated with flour and dried for durability. But at
the time of storage and marketing, the fish are often
contaminated with mold if that could reduce fish quality
wood. This is because the wooden fish is stored and sold in
traditional markets without packaging so as to enable
contamination by certain types of mold. In addition the use of
fish flour as a coating can also trigger the growth of mold
contaminants.
Based on previous research, all wood obtained from the fish
market in Banda Aceh and Lhokseumawe contaminated by
Aspergillus [1]. Besides the wooden fish sold in five tradisional
markets Banda Aceh positive note containing formaldehyde [2].
Health risks posed if consumers consume products that have
formaldehyde is very dangerous because it is a carcinogen.
Therefore, this study focused on the use of shells as a source of
chitosan in an effort to prevent mold contamination in fish
timber with reference to food grade standards.
The use of chitosan biomaterial is one alternative way to
overcome the problems of mold presence of contaminants in
fish, especially wooden molds. The advantages of using
chitosan as a coating of food is to have antimicrobial activity
and are not toxic to humans so it is safe for consumption [3],
[4], [5]. Several studies have reported that chitosan has the
ability as an antimicrobial and can be used in food [6], [7],
[8]. The use of chitosan can be applied as a coating (edible
coating) [5], [9], [10]. This study aimed to analyze the long
soaking wooden fish is optimal in solution of chitosan to
produce wooden fish that have a longer shelf life than fish
timber soaked with chitosan.
179 Resmila Dewi and Endang Sutariningsih Soetarto: The Utilization of Chitosan from Shrimp Shells as Edible
Coating for Wooden Fish (Keumamah)

2. Material and Methods
2.1. Materials
Shrimp shells as a source of chitosan, tuna as a wooden
fish, Potato Dextrose Agar, NaOH, HCl, aquadest, acetic acid.
2.2. Methods
2.2.1. Preparation of Chitosan from Shrimp
Shells
Preparation of chitosan is done by extraction, using dried
shrimp shells as raw material refined. The initial step is
deproteinasi; shrimp shell powder is placed into a vessel and
added NaOH 3.5% (1:10 w/v), then heated at a temperature of
65°C for 2 hours while stirring. Once the mixture to cool,
filtered and washed with distilled water until pH neutral and
dried result of this process is a rough chitin. The second step is
the demineralization; chitin rough weighed and added HCl 1
M (1:10 w/v), left to stand for 30 minutes at room temperature
while stirring. When finished, washed with distilled water until
pH neutral and dried at a temperature of 65°C to dry. The
outcome of this process is chitin. The third step is
deacetylation; Chitin is weighed and added 50% NaOH (1:10
w/v) in a container and stirred while heated at a temperature of
100°C for 30 minutes. After chilling filtered and washed until
pH netral and dried at a temperature of 65°C.
2.2.2. The Application of Chitosan as a
Coating on the Wooden Fish
Tuna (Euthynnus affinis) cleaned and washed with clean
water. Braised fish with salt water for ± 50 minutes and then
drained, cut lengthwise into four sections by first separating the
bones, is dried directly under the sun until the surface of the fish
becomes dry and crumble. The next fish immersed in a solution
of chitosan with long immersion for each treatment were 0, 15,
30, 45, 60 and 75 minutes. After immersion in a solution of
chitosan, drained fish. Fish that have been coated with chitosan
was dried in the sun for ±3 days until the fish become hard as
Table 1. The extraction of chitosan from shrimp shells.
wood and used as the control fish coated in flour. Furthermore,
the fish are kept at room temperature. Each specific time
intervals (once a week) changes observed through organoleptic
and fungi population analysis to determine the effect of chitosan
coating on old wooden shelf life of fish.
The storage of wooden shelf life of fish observed by
changes in color, texture, aroma and flavor of the fish
timber that has been coated with chitosan and compared
with coated wooden fish flour as a control. Organoleptic
observations made during a month on days 0, 7, 14, 21, and
28. This organoleptic test using the 5 panelists using
hedonic method (using the assessment sheet) which has a
score of 1-9 [13].
Mold on the fish population analysis of timber is made by
counting the number of colonies of mold that grows on the
wooden fish in each treatment with total plate count method. A
total of 10 g of fish samples from each treatment timber that
has been mashed suspended in 90 ml of sterile distilled water
and vorteks until homogeneous (10-1 dilution). The suspension
was taken 1 ml and suspended in 9 mL of sterile distilled water
to obtain a dilution of 10-2 and vorteks until homogeneous.
Each suspension dilution of 0.1 ml inokulation taken in the
distribution of PDA, made duplo and incubated at room
temperature for 7 days. After an incubation period completed
calculating the number of colonies that grow on the surface of
the media, is calculated using the formula:
1
total colonies
dilution factor
3. Result and Discussion
3.1. Chitosan from Shrimp Shells as a
Coating of Fish
Preparation of chitosan from shrimp shells carried by the
extraction method includes deproteination, demineralization,
and deacetylation (Table 1).

Raw material Extraction stages chitosan Result Weight (g) Yield (%)
Dried shrimp shells 8.000,0
Deproteination Chitin rough 2580.9 32.26
Demineralization Chitin 1426.4 17.83
Deacetylation Chitosan 846.1 10.57

A total of 8 kg of shrimp shells do deproteinasi process. Results obtained from chitin deprote inasi as much as 2580.8 g
coarse, demineralization as many as 1426. 4 g of chitin, and deacetylation of chitin produces 846.1 g chitosan. Color
degradation shrimp shell in chitosan extraction process can be seen in Figure 1.

Figure 1. Degradation shrimp shell color during the manufacturing process of chitosan. a) reddish brown shrimp shells; b) the results deproteination light
brown; c) demineralized (chitin) cream-colored; and d) the results of deacetylation (chitosan) cream-colored.

Chitosan is obtained and characterization include color, texture, particle size, moisture content, ash value, nitrogen
 American Journal of Food Science and Nutrition Research 2017; 4(6): 178-183 180

content, deacetylation degree and chitosan yield. Results The analysis characterization of chitosan used in this study
characterization of chitosan (Table 1). according to the standard specifications of chitosan, so that it
can be concluded that chitosan in this research can be applied
Table 2. Characteristics of chitosan. as a coating on wooden fish.
No. Specification Value Standard* [11]
1 Color Cream White to cream 3.2. Effect of Chitosan Coating on the Shelf
2 Texture Powder Powder Wooden Fish
3 Particle size 40 mesh >10 mesh
4 Moisture content 5,5% <10% The storability of wooden fish is based on the physical
5 Ash value 1,5% <2% condition of the fish through organoleptic (color, texture,
6 Nitrogen content 1,3% <5% aroma and flavor) and the presence of mold on the wooden
7 Deacetylation degree 79,74% >70%
fish during storage.
8 Chitosan yield 10,57% -

3.2.1. Organoleptic
(i). Color

Table 3. Average organoleptic value of color wooden fish during storage.

Storage times (day)

Treatment 0 7 14 21 28
organoleptic value of color
Control 9,0±0,0g 8,8±0,3fg 7,8±0,0bcd 7,2±0,3ab 7,0±0,0a
Old soaking the fish in chitosan (minute)
0 9,0±0,0g 8,8±0,3fg 7,6±0,3abc 7,6±0,3abc 7,0±0,0a
15 9,0±0,0g 9,0±0,0g 8,0±0,8cde 8,0±0,6cde 7,0±0,0a
30 9,0±0,0g 9,0±0,0g 8,4±0,3defg 8,2±0,0cdef 7,0±0,0a
45 9,0±0,0g 9,0±0,0g 8,4±0,3defg 8,2±0,0cdef 7,0±0,0a
60 9,0±0,0g 9,0±0,0g 8,6±0,6efg 8,2±0,0cdef 7,6±0,3abc
75 9,0±0,0g 9,0±0,0g 8,6±0,0efg 8,2±0,0cdef 7,6±0,8abc
SNI 01-2346-2006 Minimum 7,0

Description: a number followed by the same letter not significantly different; ± SD (n = 2; α = 0.05)

The average value of organoleptic color wooden fish in
the control treatment was not significantly different from
the old treatment of wooden fish soaking in a solution of
chitosan on all the time soaking after 28 days of storage.
However, organoleptic value of color wooden fish with a
long treatment of wooden fish soaking in a solution of
chitosan for 60 and 75 minutes better than the other
treatments after 28 days of storage. However, the average
value of organoleptic color wooden fish on all treatments
are still considered good because they conform to the
standard wooden fish organoleptic value is at least 7 (SNI
01-2346-2006) [14].

(ii). Texture

Table 4. Average values organoleptic texture of the wooden fish during storage.

Storage times (day)

Treatment 0 7 14 21 28
Organoleptic value of texture
Control 9,0±0,0b 9,0±0,0b 9,0±0,0b 8,8±0,28b 8,4±0,28a
Old soaking the fish in chitosan (minute)
0 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b 8,8±0,28b
15 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b
30 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b
45 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b
60 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b
75 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b 9,0±0,0b
SNI 01-2346-2006 Minimum 7,0

Description: a number followed by the same letter not significantly different; ± SD (n = 2; α = 0.05)

The results of organoleptic assessment texture of the texture wooden fish with flour coating treatment was not
wooden fish during storage showed that the organoleptic significantly different from the old treatment of soaking in a
181 Resmila Dewi and Endang Sutariningsih Soetarto: The Utilization of Chitosan from Shrimp Shells as Edible
Coating for Wooden Fish (Keumamah)

chitosan after 0, 7, 14, and 21 days. However, this value was
significantly different between the treatment coating flour
with long immersion in chitosan treatment after 28 days of
storage (Table 4). Tekstur a food is closely related to the
existing moisture content in these foodstuffs. The higher the
moisture content, the more soft or mushy. Wooden fish that
are still in good condition has a hard texture and not easily
broken. All the wooden fish on a soaking treatment chitosan
(iii). Aroma
has great organoleptic value. This is because the chitosan has
an amine group capable of binding water. Chitosan is one
type of polysaccharide that is a barrier is good, because the
polysaccharide coating can form a strong and compact matrix
[12]. Therefore, the chitosan coating can serve as a barrier
between materials with the environment so as to reduce the
influence of the environment on food.

Table 5. Average values organoleptic aroma of the wooden fish during storage.

Storage times (day)

Treatment 0 7 14 21 28
Organoleptic value of aroma
Control 9,0±0,0f 9,0±0,0f 8,4±0,3e 6,4±0,3b 2,5±0,4a
Old soaking the fish in chitosan (minute)
0 9,0±0,0f 9,0±0,0f 8,4±0,3e 7,4±0,6cd 7,0±0,0c
15 9,0±0,0f 9,0±0,0f 8,6±0,0ef 7,6±0,3d 7,0±0,0c
30 9,0±0,0f 9,0±0,0f 8,6±0,0ef 7,6±0,3d 7,0±0,0c
45 9,0±0,0f 9,0±0,0f 8,8±0,3ef 7,8±0,0d 7,0±0,0c
60 9,0±0,0f 9,0±0,0f 9,0±0,0f 8,4±0,3e 7,4±0,0cd
75 9,0±0,0f 9,0±0,0f 9,0±0,0f 8,4±0,3e 7,4±0,6cd
SNI 01-2346-2006 Minimum 7,0

Description: a number followed by the same letter not significantly different; ± SD (n = 2; α = 0.05)

Wooden fish on all treatments have a value of organoleptic
aromas 9 after the shelf life of 7 days. This value decreases
after a 28-day shelf life. Wooden fish soaking treatment in
chitosan for 60 and 75 minutes, giving the best results
because it has the organoleptic aromas 7.4 after 28 days of
storage. However, treatment with soaking time 0, 15, 30, and
45 minutes is still considered good because it has the
organoleptic 7 (Table 5). This value is still in accordance
with the standard wooden fish organoleptic value is at least 7
(SNI 01-2346-2006) [14].
Coated fish flour (control), the aroma is faster in a
degraded condition which is characterized by additional
odors such as rancidity. The smell arising allegedly due to
(iv). Taste
the breakdown of protein in fish. Phase splitting and damage
the protein starting from the microbial contamination of
materials. The microbes will produce a protease enzyme that
breaks down proteins into small molecular compounds [15].
The result of this solution which will ultimately affect the
smell of the food so that the aroma of the fish-covered wood
flour in this research turned into rancid. On the wooden fish
treated with chitosan coating does not cause a rancid smell
and ammonia. This is due to the chitosan coating can inhibit
the oxidation processes fat by reducing oxygen into the flesh
of fish, so the fish coated chitosan odorless. Chitosan is able
to inhibit undesirable odor by inhibiting the release of
volatile compounds that cause odor [8].

Table 6. Average values organoleptic taste of the wooden fish during storage.

Storage times (day)

Treatment 0 7 14 21 28
Organoleptic value of taste
Control 9±0.0c 9±0.0c 8,2±0.0b 8±0.3b 6.4±0.3a
Old soaking the fish in chitosan (minute)
0 9,0±0,0c 9,0±0,0c 8,6±0.1c 8,2±0,6b 7,0±0,0b
15 9,0±0,0c 9,0±0,0c 8,6±0.1c 8,2±0,0b 7,0±0,0b
30 9,0±0,0c 9,0±0,0c 8,6±0.1c 8,2±0,0b 7,2±0,3b
45 9,0±0,0c 9,0±0,0c 8,6±0.1c 8,2±0,6b 7,2±0,3b
60 9,0±0,0c 9,0±0,0c 8,6±0.1c 8,2±0,0b 7,4±0,0b
75 9,0±0,0c 9,0±0,0c 8,6±0.1c 8,2±0,0b 7,4±0,0b
SNI 01-2346-2006 Minimum 7,0

Description: a number followed by the same letter are not significantly different; ± SD (n = 2; α = 0.05)

Wooden fish soaking treatment in chitosan for 60 and 75 taste value of 7.4 after 28 days of storage. However,
minutes, giving the best results because it has organoleptic treatment with soaking time 0, 15, 30, and 45 minutes is still
 American Journal of Food Science and Nutrition Research 2017; 4(6): 178-183
considered good because they conform to the standard
wooden fish organoleptic value is at least 7 (SNI 01-2346-
2006) [14] (Table 6). Fish soaked in chitosan solution, has a
flavor that is not much different with the fish coated in flour.
This is because as edible coating of chitosan just coat the
outer surface of the fish so that chitosan does not seep into
the flesh of the fish.
Table 7. Mold population on wooden fish during storage.
Storage times (day)
Treatment 0 7
Total of mold (CFU/g)
Control - -
Old soaking the fish in chitosan (minute)
0 - - -
15 - -
30 - -
45 - -
60 - -
75 - -
SNI 01-2346-2006 Mold must be negative
Description: ±SD; n=2
The results showed that the wooden fish with chitosan
soaking treatment can reduce the rate of growth compared to
fish mold-covered wood flour. Fish-covered wood flour has
been contaminated by mold on the 14th day of storage (Table
7). The cause mold contamination allegedly due to
environmental factors. Environmental conditions with high
humidity causes damage to the wooden fish due to increased
absorption of water vapor. Water is one of the components
required by mold in the process of metabolism. Wooden fish
that have high water content accelerates the growth of mold.
Besides influenced by moisture, the use of fish flour as a
coating can also be a trigger mold growth. This is due, wheat
flour contains starch required by mold as a source of
nutrients for growth. Fungus will hydrolyze starch into
simpler compounds such as glucose, which can then be used
by the fungus to breed.
The longer storage, the amount of mold that is contained in
fish timber increased. On the shelf life of the 14th day,
treatment of wooden fish with flour already contains a
population of about 0,45×102 mold colonies/g and the value
increased at day 28 which 1,55×102 colonies/g (Table 7). It is
caused during storage, fungi have adapted to their
environment. In addition, fungi activity of cell growth
through an overhaul of the compounds contained in wood
fish such as fat and protein.
Concentration of 1.5% chitosan as a coating of wooden fish
on the storage 0-days until 21-days does not contain colonies
of mold. But the shelf life of 28 days, the wooden fish with
chitosan soaking treatment for 0, 15, 30, and 45 minutes start
to grow mold with the population of each 0,30×102; 0,20×102;
0,20×102; and 0,15×102 colonies/g, whereas the chitosan
soaking treatment for 60 and 75 minutes did not contain
colonies of mold. These results indicate that longer time the
fish soaking in chitosan solution tends the amount of mold to
182
3.2.2. Analysis of Molds Population
During storage timber all fish samples were observed
resistance to mold growth by using Total Plate Count. The
results of the analysis of the average population in the mold
of each treatment wooden fish listed in Table 7.
14 21 28
0,45×102±7,0 0,85×102±7,0 1,55×102±21,2
- 0,30×102 ±14,1
- - 0,20 ×102± 0,0
- - 0,20 ×102± 0,0
- - 0,15 ×10 2± 0,0
- - -
- - -
decrease even can not grow. Chitosan as an antifungal also
relates to its ability to coat the material. The longer the time
soaking the fish in a solution of chitosan causes more and more
powerful chitosan coating the fish so as to maintain the quality
of the fish. Chitosan has a positively charged amine group that
is capable of binding with a negative charge on the cell surface
mold that causes the expansion of the cell surface and change
the permeability of cell walls. Changes in the cell wall
permeability leading to leakage of intracellular substances such
as electrolytes, proteins, amino acids, glucose and the enzyme
that inhibits the metabolic processes that ultimately lead to
mold can not grow.
Based on the results of organoleptic test and analysis of
mold on wood fish populations, showed that soaking the fish
in a solution of chitosan for 60-75 minutes gives good results
so that it can be used as a preservative because it can extend
the shelf life of fish timber. Several studies have reported that
chitosan has the ability as an antimicrobial and can be used in
food [6], [7], [8]. Chitosan as antimicrobial coating can serve
as a secure, so that it can extend the shelf life of food [15],
[16], [17].
4. Conclusion
Based on the results of this study concluded that soaking in
a chitosan wooden fish for 60-75 minutes showed effective
results and provide good organoleptic value most, with no
evidence of mold growth after a shelf life of 28 days so that
the wooden fish suitable for consumption.
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