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.

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

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 Yield ( %) Visual appliance - Color - Odor Number of extraction Single Double Triple 28,7 Brown Weak 14,8 Brown-light No 14,2 Brown-light 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

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 : ( A1655/ A3450 x 1 /1,33 ) x 100 %, A = log P0 / P D (%) = 1-

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 Yield of chitosan from chitin Degree of deacetylation Time of deacetylation ( minutes) 120 74,4 60,4 240 72,6 65,2 300 69,6 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 .

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. Chitins 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 chitins 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. 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. analytical grade reagents from E Merck. All other chemicals NaOH, HCl were The knowledge of these containing processes is essential to optimize production technologies of chitin

The irradiation of sample for sterilization was carried out in a 2,0 kGy / hr Result of Experiment

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Co Gamma

irradiation source. The dose of irradiation were 2, 4, 6, 8, and 10 kGy with a dose rate of

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 1 2 3 4 5 6 Irradiation dose ( kGy ) 0 2 4 6 8 10 Bacteria 1,5 x 104 1,5 x 103 1,5 x 10 0 0 0 Yeast/ Mold 3 x 103 3 x 10 0 0 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

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 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. the modification technique. Graft modified of chitosan with hydrophilic functional monomers was suitable method to develop its

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 radicals method . 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. order to improve its properties such as hydrophilicity and reactivity. Graft copolymerization induced by radiation is one of the methods for polymer modification in 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, and decolorization by acetone . The irradiation of samples was carried out in
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The methanol-acetic acid in the

composition of 7 / 3 was the good solvent for grafting of chitin. The percentage of

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.

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

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 petroleumbased film or to new specialized niche areas made available by increasing because from our environmental concern. We choice oligo-chitosan as basic material for bioplastic 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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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
1.

Goosen,M.F.A. , Aplication of Chitin and Chitosan, Technomic Publishing Company, Inc, Lancaster, Pennsylvania, USA. 1997, 297 305. Purwatiningsih, Isolasi kitin dan komposisi senyawaan kimia dari limbah udang windu, Tesis .Jurs. Kimia ITB ,1992, 5 30. Kurita, K; Koyama,Y ; and Taniguchi, A. Journal of Applied Polymer Science. 1986 , 31, 1169 1176. Hong, K.N.O ; Mayers, S.P; Lee, K.S. Journal of Agricultural and Food Chemistry, 1989 , 37 ,(3) , 575 579. 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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Gatot Trimulyadi Rekso; dan N.M. Surdia, Pengaruh Dosis Total Iradiasi 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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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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