The remarkable rapid and successful expansion of prawn processing industry in Indonesia suggests the possibility of utilizing prawn-processing waste as raw material for the manufacture of many valuable products such as chitin and chitosan

1. RESEARCH AND DEVELOPMENT OF RADIATION PROCESSING OF CHITIN
AND CHITOSAN IN INDONESIA
Gatot Trimulyadi Rekso
Center for Research and Development of Isotopes and Radiation Technologies
National Nuclear Energy Agency, Jakarta, Indonesia
INTRODUCTION
Indonesia is the world's largest archipelago with around 13,000 island, of which 6.000
are inhabited, and occupies a 5,100-km stretch from the Indian to the Pacific Ocean. With
a total land area of almost 195 million hectares, it is the largest member country of the
Association of Southeast Asian Nations (ASEAN) and the most important country in
Asia regarding of ocean and lands those rich of natural polymer material.
Center for Research and Development of Isotopes and Radiation Technologies
(P3TIR) which was established in December 20, 1966 the Pasar Jumat Atomic Energy
Research Complex,
Industrial processing division is one of the several divisions in P3TIR. In this
division has five group activities, radiation polymerization, radiation sterilization and
food preservation, industrial dosimeter and waste recycle for industrial purpose. The
waste recycle for industrial purpose group is one of the new groups, which has
established in 1998. The activity of this group is research and development utilization of
shell seafood waste. For industrial purpose such as chitin and chitosan
Shell seafood waste such as prawn shell, crabs shells are abundantly found in
Indonesia. Utilization of the fishery waste mentioned above to be useful product is
suggested not only to recycle the fishery waste but also to reduce the environmental
pollution and to improve the economic situation such as to preparing of the field
employee occupation, to bring socio-economic benefit..
The remarkable rapid and successful expansion of prawn processing industry in
Indonesia suggests the possibility of utilizing prawn-processing waste as raw material for
the manufacture of many valuable products such as chitin and chitosan.
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2. The main activity of the research in waste recycle for industrial purpose is to do R & D
to provide data that will enable of shell seafood waste to be used in a commercial
production of chitin, chitosan, its derivative and its modification.
THE RESEARCH HAVE BEEN DONE
1. The isolation of chitin /chitosan from waste of fish frozen industry.
In the beginning of our research work was involved in making on the research
advisability of the natural of resource for chitin and chitosan such as various kind of
prawn and crab shell. The survey was done in side beach of north Java Island. We
found that prawn and crabs shells are easily collected from the frozen prawn
processing plant. Shell was ground using a blender. Then served to obtain particle
size of 40 + 60 mesh.
Isolation of chitosan
Dried prawn shells
Grinding
1N NaOH Deprotenization
Washing
1 N HCl Demineralization
Washing
Drying
Chitin
Figure 1. Flow diagram of the chitin preparation
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3. Chitosan can be obtained by treating chitin with 50 percent sodium hydroxide with
liquid solid ratio 20 : 1 at 1000 C for 2 hr, 3 hr and 4 hr . After this process, solids
separated from the alkali layer were extensively washed with distilled water to remove
the traces of alkali. The resulting solids were dried in vacuum oven at 500 C for 24 hr.
The preparation of chitosan can be seen in Figure 2.
Chitin solids
NaOH ,50 % Deacetylation
Drying
Chitosan
Figure2. Flow diagram of the chitosan preparation
Result of the experiment
The average yield of isolation chitin from prawn shell is shown in Table I. The data
obtained shows that the number of extraction affected the yield of chitin . It was reported
that the number of stage of extraction affected the mineral removal significantly.
Table 1. The yield of chitin at various number of extraction
Parameter Number of extraction
Single Double Triple
Yield ( %) 28,7 14,8 14,2
Visual appliance
- Color Brown Brown-light Brown-light
- Odor Weak No No
From the Table it was seen that by single stage extraction . The average yield of
chitin was 28,7 % and the solids obtained were brown and still have a weak odor. This
unexpected color and odor was caused by the presence of some protein mineral and
pigment such as carotenoid. The double one gave an average 14,8 percent and no odor,
the color was brown-light. So it is obvious that the number of extraction affected the
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4. protein and mineral removal. However for three times extraction yield of the solid was
not giving significantly different compare with double extraction. It was meaning that
double stage extraction was enough for isolation of chitin from prawn shell.
The conversion of chitin to chitosan was achieved by extracting chitin in 50 percent
sodium hydroxide with solid –liquid ratio = 1 : 20 at temperature 1000 C for 120, 240,
and 300 minutes. The data obtained gave an average yield of chitosan from chitin were
72,2 percent. Therefore the overall extraction yield of dried prawn shell in chitin form
was 14,8 percent and chitosan form was 10,6 %. The degree of deacetylation was
determined by means of FTIR spectrophotometer.It is determined by the base line
method. An infrared spectrum was recorded in a range 3500 – 500 cm -1 and absorbency
at 2870 cm-1 ( the C-H ) and 1550 cm-1 (the amide band) were evaluated by base line
method. The deacetylation degree ( D ) is calculated from equation : D (%) = 1-
( A1655/ A3450 x 1 /1,33 ) x 100 %, A = log P0 / P
Where A1655 and A3450 are absorbencies at 1655 cm-1 ands 3450 cm-1 , respectively.
Table 2. The degree of deacetylation at various time of reaction
Parameter Time of deacetylation ( minutes)
120 240 300
Yield of chitosan from chitin 74,4 72,6 69,6
Degree of deacetylation 60,4 65,2 73,0
It can be seen in Table 2. That the deacetylation time gave significant effect of
deacetylation. After 120 minutes degree of deacetylation reaches 60 4 % , it is gradually
increased as the time increased. The highest degree of the acetylation was achieved for
300 minutes.
Sehchi Mima and coworkers reported deacetylation proceed to about 70 % within the
first 1 hr of alkali treatment in 50 % NaOH solution at 100 0 C , but it progresses only
gradually after this reaching 80 % in 5 hr. Further alkali treatment does not deacetylate
significant and only degrades the molecule chain .
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5. II. Preliminary studies of chitin sterilized by irradiation
Over the past several years, chitosan has been receiving increased attention for its
applications in the chemical, biomedical, and food industries. Chitin’s key properties are
its ability to act as a cationic flocculent, viscosifier and selective chelator of metal ion.
The major commercial application for chitin currently is used for waste processing, but
there is growing interest as non-toxic cationic polymer in the hair treatment and skin care.
Clear solution form clear film that adhere to skin or hair, primary due to chitin’s cationic
character. Chitin is an excellent moisturizer, attributes
to wound healing, make an attractive biopolymer for cosmetics a personal care
application. Hun-Lee and coworkers reported for the antimicrobal activity of chitin as
natural preservatives in cosmetic product against some microorganisms . Formula
preserved with 0,5 % chitosan had an effective antimicrobial activity against the Gram
(+) and Gram (-) bacteria but not fungi.
In order to develop application of chitin as cosmetic additive it is necessary to choose
the best way of its sterilization. Chitin easily undergoes thermal destruction, chemical
sterilization by ethylene oxide or formaldehyde was proved to reduce its
biocompatibility. On the other hand, the radiation sterilization leads even to improvement
of some biocompatibility factors.
The aim of our work was to determine of irradiation sterilization dose of chitin from
shell waste of prawn (penaeus monodon). Analysis of change in infrared spectra and
thermal stability before and after irradiation also studied. The knowledge of these
processes is essential to optimize production technologies of chitin containing
biopolimer used as additive for cosmetic.
Material and Experiment
Chitin extracted from prawn shell ( Penaeus monodon ), it was got from Muara Karang ,
North Jakarta. They were initially washed by water and then dried at 800 C overnight and
conditioned at room temperature for 24 hr. All other chemicals NaOH, HCl were
analytical grade reagents from E Merck.
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6. 60
The irradiation of sample for sterilization was carried out in a Co – Gamma
irradiation source. The dose of irradiation were 2, 4, 6, 8, and 10 kGy with a dose rate of
2,0 kGy / hr
Result of Experiment
Results concerning the effect of radiation sterilization by various doses on the
total number of bacteria of chitin are presented in Table 3.
Table 3. Total number of bacteria
No Irradiation dose Bacteria Yeast/ Mold
( kGy )
1 0 1,5 x 104 3 x 103
2 2 1,5 x 103 3 x 10
3 4 1,5 x 10 0
4 6 0 0
5 8 0 0
6 10 0 0
It was found that at a dose of 4,0 kGy the number of yeast and mold was eliminated
and for bacteria was at 6,0 kGy. On the basic of the above result, it shows that the dose
of irradiation sterilization was not so high . So it was irradiation seems to be best
sterilization method for sterilized of chitin.
III. Studies on radiation graft-copolymerization of monomer with selective
functional group onto chitin as adsorbent.
The natural chelating marine polymer chitin, Polly(N-acetyl-D-glucosamine) and its
deacetylated derivative chitosan are useful for removing heavy metal ion waste from
discharge water . Chitin , the most abundant naturally is undoubtedly one of the most
promising and attracting resources present in quantity. Among some interesting
properties of chitin , chelating ability arising from its characteristic structure is especially
noteworthy. Many researchers have explored the feasibility of this approach. Kurita.K
et.,all, conducted experiments with a number of heavy metals.
The adsorption behavior of chitin and its concerns with various degree of
deactylization. This high adsorption capacity was ascribable primarily to its remarkable
hydrophilicity in cooperation with the relatively high amino group content. It is indicate
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7. that the importance of hydrophlicity and suggest that , in order to develop adsorbents of
high capacity , it is make indicate the importance of hydrophilicity essential to make
chitin derivatives highly hydrophilic and yet insoluble in water.
Chitosan is natural polysaccharide and has the same skeleton structure as
cellulose, by a radiation modification such as graft-copolymerization of
hydrophilic monomer expected to improve its hidrophilicity.and performance for
application as an ion exchange adsorbent. In the field of Radiation Process, free
radical formation is the key role of the modification technique. Graft modified of
chitosan with hydrophilic functional monomers was suitable method to develop its
as ion exchange adsorbent
The experiment have been done were list above :
A. The effect of total irradiation dose on graft copolymerization of acrylic acid onto
chitin .
The effect of total irradiation dose on graft copolymerization of acrylic acid onto
chitin by pre-irradiation technique at nitrogen atmosphere has been carried out. The aim
of this research was to find out the optimum of total dose of grafting with water as
solvent at various times of reaction and temperature. Instead of that the effect of total
dose on the molecular weight and the free radicals formation were evaluated. The
influence of monomer concentration and solvent composition were studied at optimum
total dose. The result showed that the percentage of grafting increased with increasing of
the total dose and the optimum condition was achieved at the total dose of 12 kGy . The
solvent composition has significant effect on the degree of grafting . The best
composition was obtained in the acetic acid /methanol weight ratio 1 : 1. The percentage
of grafting increased with increasing the monomer concentration. Visual observation
showed that the formation of homopolymer was affected by the reaction temperature and
concentration of monomer.
B. The effect of solvent composition on grafting of acrylic acid onto chitin by
irradiation technique.
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8. The effect of solvent composition on grafting of acrylic acid onto chitin by irradiation
technique have been carried out. The aim of the research is to find out the optimum
solvent composition to the increase swelling properties of chitin so that acrylic acid
could carry in the active site of chitin. In this experiment the trapped radical and
peroxide graft irradiation co-polymerization method were employed . The influence of
solvent compositions e.g. water – methanol, water – acetic acid, and methanol –acetic
acid and the grafting method that affected to the degree of grafting have been studied.
The result showed that the percentage of grafted chitin by pre- irradiation peroxide
method is higher than trapped radical’s method . The methanol-acetic acid in the
composition of 7 / 3 was the good solvent for grafting of chitin. The percentage of
grafting was found 47,2 %.
C. Study on radiation grafting of acrylamida onto chitin
Studies on radiation grafting of chitin were examined. Chitin is amino
polysaccharides and thus anticipated to have high potential as specialty polymeric
materials compared with cellulose. They are, however, utilized scarcely because of
problems associated with poor solubility, hidrophilicity , and low reactivity. Graft
copolymerization induced by radiation is one of the methods for polymer modification in
order to improve its properties such as hydrophilicity and reactivity. In this present
study , modified chitin was prepared by grafting of acrylamide using gamma ray. The
grafted functional group onto chitin are expected to increase its hydrophilicity and
reactivity.
Isolation of chitin involved basic operations such as deproteinization, demineralization,
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and decolorization by acetone . The irradiation of samples was carried out in Co
gamma irradiation source . In the present experiment, the pre- radiation –peroxidized
method was employed. In this method chitin was irradiated in presence of atmospheric
oxygen at room temperature. A monomer acrylamide solution than introduced into
irradiated chitin and the graft polymerization was carried out in a nitrogen atmosphere at
a certain time of reaction. The grafted chitin obtained was washed and dried in vacuum
and the percentage of grafting was measured by gravimetric method.
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9. The percentage of grafting has been determined as a variation of total dose, monomer
concentration and temperature as a function of time of reaction. The results showed the
optimal total dose for the grafting process was 12 kGy. The percentage of grafting
increases as increasing total dose, acrylamide concentration and temperature. The
optimal conditions were total dose of 12 kGy. acrylamide concentration of 30 %,
temperature of 70 0C and reaction period of 3 hours. The yield of grafting was found
94,3 %. The presence of acrylamide on chitin was demonstrated by IR spectrum and the
thermal stability by DSC.
THE RESEARCH STILL BE DOING
I. Removal of heavy metals from solution by using modified chitosan (Chito-g-Aac )
Selective removal of heavy metal such as Cu, Hg, Cd and Pb from solution at
laboratory scale was carried out. The most promising method of removing of heavy
metal from solution is the use of a cation –exchange materials having a chelating group
such as Chitos-g-Aac or Chitos-g-Aam.. A previous study in our laboratory has
indicated that Cu ion can be successfully remove from the solution using chitosan
modified with value of 8 times compare only non modified chitosan. The goal of the
study was to investigate the possibility of removing toxic heavy metal such as Hg, Cd,
and Pb
II. Study of composition on formulation of chitosan as film forming for nail polish
The effect of solvent composition on formulation of chitosan as film forming has been
carried out. The aim of this research was to find out the best composition of solvent using
chitosan as film forming for nail polish. The solvent used were ethyl alcohol, butyl acetic
acid, and acetic acid. The physical and mechanical properties of the resulting film were
examined, e.g. hardness, adhesion and glossy of the film, viscosity and flow properties of
the formulations.
The results shows that the best formulation for chitosan as film forming were as follows;
concentration of chitosan 2 %, the composition of the solvent was ethyl alcohol / acetic
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10. acid = 1 / 3 with the value of the hardness of the film B, adhesion 78,5 %, glossy 68,2 %
and the viscosity of the formulation was 1113 cps with a good flow properties.
In this research we continue study of effect an organic pigment and some additive like
hardener etc for make a good film for nail polish .
THE RESEACH WILL BE DOING
1. Science and technology for rural areas development
This research will begin next year it was our division proposal to our government for
developing of rural people in north sea side Java Island , which that area rich with shell
shrimp or crab waste. In that proposal we will introduce and teach them how to make
chitin from that waste after that they can make by themselves and sells the product to
private company as distributor of chitin.
Chitin from distributor was send to our institute for irradiation and we process to
make oligo-chitosan with high degree of deacetylization and we send to distributor again
for distribute oligo-chitosan to industry in Indonesia. For this project we begin with the
capacity of 1000 kg chitin/ month / area, and we plans for four area there are 2 in
Cirebon and 2 in Semarang.
2. Research and development of oligo-chitosan for bioplastic materials.
One objective of research is to make biodegradable films from renewable sources
such as oligo-chitosan. These films can then be applied to replace some petroleum-
based film or to new specialized niche areas made available by increasing
environmental concern.
We choice oligo-chitosan as basic material for bioplastic because from our
experience research on chitosan as film forming we found that oligo-chitosan can
form a good film. Casting on glass plate at elevated temperature can form Chitosan
films. For getting a good film some additive must be added on the formulation such
as crosslingking agents and plastisizer.
CONCLUSION
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11. ∗ It is clear that the research give highlights the probability of utilizing. The waste
product of the sea food industry (prawn, crabs shell) to products which are useful in a
number of different area.
∗ Irradiation treatment was applicable on upgrading of the chitosan. Such as
sterilization, degradation to make oligo-chitosan and modification of chitin by
irradiation grafting.
REFERENCES
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Inc, Lancaster, Pennsylvania, USA. 1997, 297 –305.
2. Purwatiningsih, “ Isolasi kitin dan komposisi senyawaan kimia dari limbah udang
windu”, Tesis .Jurs. Kimia – ITB ,1992, 5 – 30.
3. Kurita, K; Koyama,Y ; and Taniguchi, A. Journal of Applied Polymer Science.
1986 , 31, 1169 – 1176.
4. Hong, K.N.O ; Mayers, S.P; Lee, K.S. Journal of Agricultural and Food Chemistry,
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5. Gatot Trimulyadi Rekso; Anik sunarni; Kadariah; Isni Marliyanti; and Rahayuningsih
Chosdu, Preliminary Studies of Chitin Sterilized by Irradiation, Proceedings book of
4th Scientific Conference of The Asian Societies of Cosmetic Scientists, Bali,
Indonesia, 7 - 9 April ,1999.
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Kopolimerisasi Cangkok Asam Akrilat Pada Khitin, Prosiding Temu Ilmiah Jaringan
Kerjasama Kimia Indonesia Seminar Nasional VIII Kimia dalam Industri dan
Lingkungan, Hotel Santika Yogyakarta, 16-17 November,1999.
7. Gatot Trimulyadi Rekso; dan N.M. Surdia; Pengaruh Pelarut Pada Kopolimerisasi
Cangkok Asam Akrilat Pada Khitin Dengan Teknik Iradiasi, Seminar Kimia Bersama
ITB-UKM IV, Yogyakarta, 9-10 Februari ,1999.
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12. 8. Dewi Wulandari, dan Gatot T.M, Pengaruh Iradiasi Kobalt-60 Terhadap Khitosan
Yang diperoleh dari Isolasi Kulit Kepala dan Badan udang putuh, Tesis Universitas
Pancasila Fakultas Farmasi, Jakarta, 1999.
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