Characterization of chitin and chitosan

synthesized from red snapper (Lutjanus sp.)

scale’s waste
Cite as: AIP Conference Proceedings 1862, 030108 (2017); https://doi.org/10.1063/1.4991212
Published Online: 10 July 2017

N. D. Takarina, and A. A. Fanani

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AIP Conference Proceedings 1862, 030108 (2017); https://doi.org/10.1063/1.4991212 1862, 030108

© 2017 Author(s).
 Characterization of Chitin and Chitosan Synthesized from
Red Snapper (Lutjanus sp.) Scale's Waste
N. D. Takarina1, a) and A. A. Fanani2
1
Departement of Biology, Faculty of Mathematics and Natural Sciences (FMIPA),
Universitas Indonesia, Depok 16424, Indonesia
2
Marine Postgraduate Study Program, Department of Biology,
Faculty of Mathematics and Natural Sciences (FMIPA), Universitas Indonesia, Depok 16424, Indonesia
a)
Corresponding author: veri641@gmail.com

Abstract. Chitin and chitosan are natural biopolymer which are useful for industrial, medical and environmental field.
Study about using fish scale sources especially saltwater fish is still limited. Red snapper (Lutjanus sp) is common
tropical saltwater fish that known as important source of marine products, particularly in Indonesia. Correspondingly, the
consumption of this species has generated significant amount of discarded scale wastes recently and hence can cause
adverse impact on the environment. Utilizing the fish scale as alternative sources of chitin and chitosan can be one
solution dealing with environmental problem. Therefore, this research aimed to characterize the chitin and chitosan
derived from the red snapper scale wastes. Chitin were extracted by deproteination and demineralization while chitosan
using deacetylation. Morphology of the chitin and chitosan were analyzed using electron dispersal spectroscopy (EDS)
and scanning electron microscope (SEM), while degree of deacetylation using fourier transform infrared spectroscopy
(FTIR). Proximate analysis showed that content of moisture, ash, and nitrogen in chitin were 3.20 %, 2.40 %, 0.04 %,
respectively while in chitosan were 6.14 %, 1.18 %, 0.03 % respectively. Furthermore, amount of C, O, Na, Al, P and Ca
elements were obtained from chitin and chitosan samples by energy dispersed spectroscopy respectively. The degree of
deacetylation for both chitin and chitosan showed high value more than 75 %. Hence, by considering the chemical
properties of red snapper scales, it confirms that this species is a promising alternative source for both chitin and
chitosan.

INTRODUCTION
Chitin usually extracted from shrimp and crab shell. This compound also can be extracted from insect like
silkworm [1]. Derivative of chitin is chitosan. This biopolymer can act as a pollutant reductor and also degradable
compound [1].
Beside from crab and shrimp shell, some studies showed that chitin and chitosan can be extracted from
freshwater fishes. Zaku et al. [2] succeeded in isolating chitin from common carp (Cyprinus carpio) scale.
Moreover, Uawonggul et al. [3] and Boarin-Alcalde and Graciano-Fonseca [4] managed to extract chitin and
chitosan from Nile tilapia (Tilapia nilotica) scale. Kumaria and Rath [5] stated that chitin and chitosan extracted
from rohu (Labeo rohita) scale, and Muslim et al [6] performed extraction of chitosan and carboxymethyl chitosan
from same fish.
Chitosan from marine resources was mostly obtained from crab or microorganism. Information about chitin and
chitosan extracted from saltwater fishes is rare. This study is aimed to investigate the extraction and characterization
method in saltwater fish especially red snapper (Lutjanus sp.).

International Symposium on Current Progress in Mathematics and Sciences 2016 (ISCPMS 2016)
AIP Conf. Proc. 1862, 030108-1–030108-5; doi: 10.1063/1.4991212
Published by AIP Publishing. 978-0-7354-1536-2/$30.00

030108-1
 RESEARCH METHODOLOGY

Raw and Chemical Material

The fish scale of red snaper were bought from PT. Bening Jati Anugrah, Bogor, West Java. Scales were washed
with running water to remove dirt and other objects. Scales then dried for one day to dry. This was done at
Laboratorium Pengolahan Hasil Perikanan, Balai Besar Pengujian Penerapan Hasil Perikanan, Jakarta. Chemical
used were Sodium hydroxide (concentration 4.2 % and 80 %) and hydrochloric acid (molarity 2 N).

Research Procedure
Extraction of chitin and chitosan were carried out at Laboratorium Pengolahan Hasil Perikanan, Balai Besar
Pengujian Penerapan Hasil Perikanan, Jakarta. Chitin was obtained after through two phases, deproteinization, and
demineralization. Deproteinization was carried out by solving fish scale into NaOH solution with comparation 1:6.
Next, the mixture was heated for 5 hours in 60 °C then stirred. The mixture was filtered and residue was washed
using aquadest. Next step was demineralization. This step was done by solving the residue using HCl, solution for 6
hours with the same ratio. The mixture was treated with the same way until the residue has neutral pH.
Chitosan is obtained from chitin through deacetylation. This step is done by solving chitin using NaOH 80 % for
4 hours at 110 °C with ratio 1:3. The mixture then treated with the same way until the pH goes neutral.

Characterization of Chitin and Chitosan

Characterization of chitin and chitosan from red snapper fish scales were done in accordance with [7] and [8].
Some parameters measured were moisture content (%), ash content (%), nitrogen content (%) and the degree of
deacetylation. This analysis was done at Laboratorium Saraswanti Indo Genetech.
Moisture and ash content were measured using oven method. Moisture content was calculated using Equation 1,
while ash content was measured using Equation 2.

W 3 − W1
% moisture = × 100% (1)
W 2 − W1
with W3 is the weight of dried chitosan and Petri dish, W2 is the weight of chitosan and Petri dish, and W1 is the
weight of Petri dish only.

W 3 − W1
% ash = × 100% (2)
W 2 − W1
where, W3 is the weight of chitosan ash and Petri dish, W2 is the weight of chitosan and Petri dish, and W1 is the
weight of Petri dish only.
Nitrogen content is analysed using Kjedahl method. Content is calculated using equation 3.

(Va − Vb) xNx14.700

%N = × 100% (3)
Wx1000
Where Va is sample titration volume of HCl , Vb is void sample volume of HCl, N is normality of HCl, and W is
chitosan weight.
The degree of deacetylation calculated by displaying the infrared light on a chitin and chitosan samples then the
infrared absorption was recorded. This analysis was performed using fourier transform infrared spectroscopy at
Laboratorium Biofarmaka IPB, Bogor. Hydroxyl groups were calculated at a wavelength of 3450 cm-1, while amide
group at a wavelength of 1655 cm-1. The degree of deacetylation of chitosan is determined based on Domszy and
Roberts [9]:

030108-2
 A1655 100
% DDA = 100 − ( × ) (4)
A3450 1.33
where, A 1655 was chitosan absorbance at the 1655/cm wavelength and A 3450 was chitosan absorbance at the
3450/cm wavelength.
The microstructure of chitin and chitosan were observed using scanning electron microscope (SEM) and
equipped with energy dispersive spectroscopy (EDS). This observation was done at BATAN Puspitek Tangerang.

RESULTS AND DISCUSSION

Proximate Analysis
Proximate analysis is series analysis to obtain some values like moisture content, ash content and nitrogen
content. The eesult of this analysis due to chitin and chitosan was shown in Table 1. Based on this analysis, chitin
and chitosan extracted from red snapper were in accordance with standard set by [7] and [8]. However, this was
contrast due to nitrogen content. Deacetylation process only eliminate the acetyl group from chitin and increased
number of amin group on chitosan.

Degree of Deacetylation
The degree of deacetylation is determined based on the absorption wavelength which was 3450 cm-1 for
hydroxyl and 1655 cm-1 for amide group respectively. Absorption of wavelength is calculated based on FTIR graph
absorbance obtained from chitin (Fig. 1a) and chitosan (Fig. 1b). Based on FTIR graph, it can be seen that the
degree of deacetylation from chitin was 73.60 %, while the degree of deacetylation from chitosan was 90.83 %.

Microstructure of Chitin and Chitosan

Characterization of chitin and chitosan using scanning electron microscope was done on the magnification of
500 x (Fig. 2). Based on SEM characterization it is known that deproteinization and demineralization steps show the
effect on chitin produced as well as deacetylation step to chitosan.
Figure 2a showed that chitin from red snapper fish scale have multiple smooth layer with fracture and scar.
However, it was contrast to Fig. 2b, which showed random, unregular, unsmooth layer of chitosan. Percentage of
element content observed in chitin and chitosan were showed in Table 2. Based on Table 2, oxygen and calcium
have highest content both on chitin and chitosan.

(a) (b)
FIGURE 1. FTIR graph of (a) chitin and (b) chitosan

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 TABLE 1. Content of moisture, ash, and nitrogen on chitin and chitosan
Test result SNI Standard
Parameters Chitin Chitosan Chitin Chitosan

Moisture content 3.20 2.88 Max 12 Max 12

Ash content 2.40 1.10 Max 5 Max 5
Nitrogen content 0.04 0.01 Max 5 Max 5

TABLE 2. Element content in chitin and chitosan

Percentage (%)
Material
Carbon Oxygen Natrium Aluminium Phospor Chlorid Calcium
chitin 7.49 47.12 0.50 0.28 15.76 0.31 28.54
chitosan 10.25 45.65 2.31 0.77 13.58 - 27.44

(a) (b)
FIGURE 2. Characterization of (a) chitin and (b) chitosan using SEM

(a) (b)
FIGURE 3. The energy level of (a) chitin and (b) chitosan resulted from EDS.

The energy level of chitin and chitosan were performed in Fig. 3. Figure 3a and 3b showed that chitin and
chitosan did not contain hazardous metal. The existence of phosphorus (P) and chlorine (Cl) in scale, in line with the
statement of [10] who stated that scales also have characteristics that are found in other structures such as bones,
teeth, and tendons.

030108-4
 CONCLUSIONS
Red snapper fish scales has potential as an alternative source of chitin and chitosan. Chitosan red snapper fish
scales have a high degree of deacetylation up to 90.83 %.

ACKNOWLEDGMENTS
We acknowledge Ministry of Research, Technology and Higher Education and Directorate of Research and
Community Engagement, Universitas Indonesia for its funding through Grant of PITTA with contract number
2047/UN2.R12/HKP.05.00/2016.

REFERENCES
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7. National Standardization Agency of Indonesia, SNI 7948:2013 (24 December 2013).
8. National Standardization Agency of Indonesia, SNI 7949:2013 (24 December 2013).
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Processing Steam,” in Maximizing the Value of Marine by Products, edited by F. Sahidi (CRC Press, Bocca
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