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Indonesian Journal of Science and Education Volume 3, Number 1, April 2019, pp: 1 ~ 9 p-ISSN: 2598-5213, e-ISSN: 2598-5205, DOI: 10.31002/ijose.v3i1.911 e-mail: [email protected], website: jurnal.untidar.ac.id/index.php/ijose

The Usage of Chitosan from Shrimp Waste as Natural Preservative for Fish Cilok (Traditional Food in )

H.B.A. Jayawardanaa), Ahmad Afandib) IKIP PGRI Jember, Jln.Jawa No.10, Jember, Indonesia, Telp (0331) 335823 e-mail: a)[email protected], b)[email protected]

th th th Received: September 13 , 2018 Revised: October 23 , 2018 Accepted: March 15 , 2019

ABSTRACT Cilok is one of the traditional food in Indonesia. Cilok is a popular treat among Indonesians due to its good taste and cheap price. To make it last longer, cilok is sometimes added with preservative. However, such attempt will endanger consumers if dangerous preservatives are used, such as formaldehyde and borax. Their usage in food preservation may negatively impact consumers’ health. The aim of this research was to develop an alternative, natural preservative for fish cilok. The preservative would be chitosan. The method used is an experiment that is by making a chitosan solution from the shrimp skin waste. In the experiment, chitosan was extracted from shrimp waste. A 1,5% chitosan concentrate was used. Its preservation potency was tested by giving three treatments towards fish cilok: fish cilok with 1,5% chitosan coating; fish cilok mixed with 1,5% chitosan solution and fish cilok without chitosan. The three treatments were observed and an organoleptic test was performed according to the preservation time in a room temperature on Day 1, 2, 3, 4 and 5. The parameters tested during the experiment were taste, color, odor and texture of fish cilok. The research results showed that fish cilok coated with 1,5% chitosan solution lasted longer than fish cilok without or added with chitosan.

Keywords: chitosan, natural preservative, fish cilok

INTRODUCTION borax or formaldehyde which are dangerous to human health. Cilok is one of popular treats in Cilok with borax as its preservative Indonesian community. It is children’s may endanger children’s health. According favourite; one can find them easily in to Safira (2014), one of children’s sideroads or schools. According to favourite is cilok. Children love its Yanuarlan (2015), the term cilok is an because it has delightful shape and chewy acronym of Sundanese words aci dicolok texture. One needs to be careful when (skewered aci), a meatball shaped food buying cilok in roadside vendors because made from starch (aci) and served some of them might add dangerous food using wood stick/. additive to cilok they sell. Cilok is not only made from tapioca Side effects resulted from consuming starch, but also mixed with beef, chicken food which contains borax are irritations in or fish meat. It is a favourite treat for both digestion canals, shell and eyes, adults and children, since it is relatively stomachache, headache, severe pain in cheap and easy to find. Fish cilok tastes upper part of stomach. Borax consumption best, but decays quickly. In order to make for a long period may result in liver it last longer, some fish cilok makers or disease, body circulation system failure vendors use synthetic preservatives such as and death. Consuming 5-10 gram borax may result in shock and death on children.

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Several misuses of borax are applied on shrimp shells, reducing the environmental foods such as meatballs, cilok, contamination loads (Agustina et. al., (rice cake) and gendar cracker (Deviana, 2015). 2015). Chitosan is an organic compound Food preservatives should be made derived from chitin. It is polyelectrolyte, from natural substances which do not non-poisonous, and biodegradable. Its produce negative side effects on health. hydroxyl and amine groups make it Therefore, new innovations in producing suitable as preservative and color natural food preservatives should be stabilizer. Chitosan can be used as a attempted. One of them is the usage of preservative since it can slow down the chitosan. Chitosan is made from shrimp growth of destructor microorganisms. waste, especially shell and head parts Chitosan can be coated on food, because they contain chitin. minimizing the interaction between the Chitosan is widely used in industries as product and its surrounding environment a preservative alternative to formaldehyde (Soegiarto et. al., 2013). and borax. Preservatives are additional According to a study by Killay (2013), substances added in food to reduce chitosan could be used as an anti-bacteria activities of microorganism, slowing down additive in comestibles. Such finding is the decay process. Chitosan contains supported by Hafdani (2011) who argues positively charged polycations which are that chitosan is antimicrobial as it could able to reduce the growth of bacteria and delay the growth of phatogenic bacteria molds (Harjanti, 2014). and decomposing microorganisms, Chitosan can be derived from chitin including fungi. contained in shrimp shell. Shrimp shell Chitosan could also be used as a contains beneficial nutritions such as preservative for fish. Fish decays rather proteins, calcium and chitin. Proteins and quickly. The decay process is caused by calcium can be used as additives in animal activities of enzymes, microorganisms and feed. Shrimp waste used in this research oxidation within fish body. A spoiled fish were head and shell parts which cover is indicated by unpleasantly sharp smell, 35%-50% total weight of shrimp. inelastic flesh, cloudy eyes and sticky According to Marganov (2003), shrimp slime in gills and body (Arifin & Nugroho, shell contains proteins (25%-40%), 2016). calcium carbonate (45%-50%) and chitin A good way for preserving fish is to (15%-30%). Taolee (in Agustina, 2015) soak it in water mixed with 1.5% chitosan. explains that chitosan is a derivation from Another is to spray fish with 2.5% chitin with structure of [β-(1-4)-2-amine - chitosan. Such attempts may prolong fish 2-dioxide-Dglucose], a result of preservation for another 5 hours (Silvia et. deacetylated chitin. al., 2014). Chitosan is a polycationic polymer. Its Chitosan can also be used to preserve hydroxyl and amine groups in the polimer fruits. A study by Trisnawati et.al. (2013) chain allow chitosan to effectively bind shows that chitosan can be used to heavy metal cations as well as cations preserve lanzones through coating. from proteins and fats. Chitosan can also Coating can be done by dissolving form an absorbing membrane during the chitosan powder in acetic acid solution. binding of organic and anorganic Next, soak fruits into the mix. Coating compounds. These make chitosan more may reduce the loss of water, oxigen, useful than chitin (Sanjaya et. al., 2007). flavour and dissolved ingredients in As chitosan possesses a high economic several products, making it effective in value, therefore it is important to conduct a preserving food quality. research for producing chitosan from

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Helander (2001) states that chitosan effectiveness as a preservative. It was compound disrupts outer membranes of conducted during May – November 2018. gram-negative bacteria. Chitosan This research needs to be done because to compound, as an antimicrobial agent, can determine the effectiveness of chitosan as be added to comestibles since it is safe for a natural preservative of cilok fish. The human. Chitosan cannot be digested by tools and materials used are presented in human therefore it does not have calorie the following table. values and is discharged with feces. A factor influential on the capability of Table 1. The Tools and Materials for Making chitosan as a preservative is the number of Chitosan and Fish Cilok amine groups it contains. The number of Name Tools Materials Chitosan Bunsen burner, Fish heads and amine group depends on the removed tripod, clamps, shells, acetate acetyl group. The removed acetyl group is beaker glass, acid, HCI 1N, called the degree of deacetylation. measuring cup, NaOH 3,5%, Therefore, the higher the degree of volumetric flask, aquadest, deacetylation is, the better the capability of stirrer, NaOH 50%, thermometer, and and methanol. chitosan as a preservative will be. The litmus paper. application of chitosan as a preservative is Fish Cilok Knife, cutting Fish fillets, gained in the best condition of the degree board, basin, tapioca flour, of deacetylation (70.34%). The best blender, spoon, starch, wheat, soaking time is 45 minutes in 2% chitosan mortar, pestle, sugar, salt, pan dan stove pepper, (Harjanti, 2014). dan water. Chitosan has several benefits such as possessing a structure familiar to cellulose A Procedure for Producing Chitosan viber contained in fruits and vegetables. Chitosan is produced from a chitin Various hypotheses regarding the working isolation process. Chitin can be gained mechanism of chitosan as an anti-bacterial from a chitin isolation process from agent address its affinity which is strong shrimp shells, consisting of two steps, with microbes’ DNA, allowing it to bond demineralisation and deproteinisation. wih DNA and then disrupt mRNA and Demineralisation is conducted to eliminate protein synthesis (Killay, 2013). organic compounds cointained in the Therefore, a further research on the usage shrimp waste. Shrimp shell generally of chitosa as a natural preservative is contains 30-50% minerals (dry weight). needed. The aim of this research was to Dominant minerals found in shrimp are describe the creation process of chitosan calcium in form of CaCO3 and a small and its effectiveness as a natural amount of Ca3(PO4)3. A scheme of preservative for fish cilok. chitosan extraction is presented in Figure 1. RESEARCH METHOD Demineralisation can be conducted by adding HCI 1 N with the 1:7 ratio of the This research is experimental, weight of materials and the volume of conducted in a laboratory for producing extractor for one hour in 90ºC temperature. chitosan from shrimp waste and creating Deproteinisation aims to eliminate proteins fish cilok. Fish cilok is added with chitosan from shrimp waste. Its effectiveness in order to discover effectiveness of depends on the power of the alkaline chitosan as a natural preservative for fish solution and the degree of temperature cilok. used. A 3,5% NaOH solution heated in This research was conducted in a 90ºC temperature for one hour can be used laboratory of FMIPA IKIP PGRI Jember as an alternative to deproteinisation with a for producing chitosan and test its

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1:10 ratio between dry shrimp waste and solution (Suptijah et. al.,1992). Fresh fish

Sorting Shrimp heads/shells Cleaning Washing Filletting Drying Washing and drying Blending

Geting rid of fishy smell Demineralisati 90ºC, 1 hour on Smoothening fillets Filtering, Washing

Deproteinisati 90ºC, 1 hour Adding seasoning on Filtering, Washing Mixing with cilok dough

Deacetylatio 120-140ºC, 1 Fish cilok n hour Washing Figure 2. A procedure for Making Fish Cilok Drying (Source: Suprapti, 2003)

A Procedure for Testing Chitosan as a

Figure 1. A Procedure for Producing Chitosan Food Preservative (Source: Suptijah, et. al., 1992) In this research, a test on chitosan as the fish cilok preservative was conducted Chitosan is produced by eliminating by giving three treatments as presented in acetyl group (-COCH3) from chitin by Table 2. using a NaOH concentrated solution (50%) with a 1:20 ratio for one hour in 120- Table 2. A Procedure of Chitosan Test 140ºC temperature (Suptijah et. al., 1992). Treat Notes Observation The more acetyl groups are eliminated ment from the chitin polymer, the stronger the 1 Fish cilok were boiled, Day: 1, 2, 3, dried and dipped into 4, 5 interaction among ions and hydrogen 1,5% chitosan solution bonds from chitosan will be (Suptijah et. for two minutes. After al., 1992). that, they were dried and preserved in a A Procedure for Making Fish Cilok room temperature. 2 Before processed, fish Day: 1, 2, 3, According to Suprapti (2003), a cilok were mixed with 4, 5 general overview of making fish cilok is a 15 cc of 1,5% presented in Figure 2. chitosan solution. Fish fillets, after 5 minutes untouched, After that, they were are blended until smooth and put them into boiled, dried and preserved in a room a bowl. Mix them with ingredients, temperature. whipped egg whites and tapioca flour until 3 Fish cilok were boiled, Day: 1, 2, 3, a homogeneous, shapeable dough is dried and preserved in 4, 5 formed. Make small balls (cilok) from the a room temperature dough. Then, boil cilok until they are without chitosan. floating. After that, drain them.

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Data Collection and Analysis into an amine group (-NH2) with the Techniques addition of strong bases such as NaOH. Research data were collected via The deacetylation reaction of chitin is documentation and organoleptic tests basically an amide hydrolysis reaction of (Harjanti, 2014). Documentation was done α- (1-4) -2-acetamide-2-deoxy-D-glucose. by taking photos of fish cilok during the The OH-ion concentration is very experiment. The organoleptic tests which influential on the process of releasing the were performed consisted of specifications acetyl group from the chitin acetamide on the whole appearance such as taste, group. In this process, the powder was colour, odor and texture. The tests were mixed with 6 litres of 50% concentrated conducted towards the three treatments: 1) NaOH. During the chitosan creation the fish cilok with 15% chitosan coating; process, the amount of chitin produced the fish cilok mixed with 1,5% chitosan was 50 gram while the amount of chitosan and 3) the fish cilok without chitosan. produced was 30 gram. Therefore, the Observations on each treatment had been rendemen in the chitosan creation was done for 5 days consecutively. 10%.

RESULTS AND DISCUSSION Fish Cilok The type of fish used for making cilok Chitosan was Treadfin Bream (Nemipterus In this research, shrimp heads and nematophorus). In this research, the dough shells were used to produce chitosan. was mixed with 1 kg fish fillets. Next, the heads and shells were dough was divided into three smaller blended, resulting in shrimp shell powder. portions. Cilok which is made of three The amount of the shrimp heads and shells parts has the same color, smell, texture and was 300 gram while the amount of HCI taste, namely white, chewy texture, and 1N used in the demineralisation was 2,1 fishy taste. liters. Small balls (cilok) from the three The demineralisation resulted in pink smaller portions with 3 cm diagram were shrimp head and shell powder. The powder shaped. The first portion was used to then was used in the next phase, create cilok with 1,5% chitosan coating. deproteinisation. The deproteination phase The second portion was used to create is carried out by reacting the result of cilok mixed with 1,5% chitosan and the demineralization with strong base (NaOH), last portion was used to create cilok so that the protein will dissolve in NaOH without chitosan. solution. This deproteination reaction aims Cilok fish in the first treatment, after to break the bonds between protein and cooking then draining and waiting until the chitin by adding sodium hydroxide. In this temperature is not too hot. Furthermore, process, the powder was mixed with 3 fish cilok is dipped in 1.5% chitosan litres of 3,5% NaOH. solution for 2 minutes. The fish is then The deproteinisation resulted in white drained and stored at room temperature. In shrimp head and shell powder. The powder this first treatment, chitosan protects cilok was then used in the next phase, fish from the outer surface of the cilok, so deacetylation. According to Dompeipen, it can protect the cilok from oxidation and et.al (2016), Isolation of chitosan protect it from bacterial contamination. compounds was obtained by performing a Cilok fish in the second treatment, deacetylation reaction on chitin. before cilok is cooked, cilok is mixed with Deacetylation is the process of converting a 1.5% chitosan solution of 55 cc. Cilok an acetyl group (-NHCOCH3) to chitin then cooked. After cooking, cilok is drained and stored at room temperature. In

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this second treatment, the possibility of and minimize any contaminant caused by chitosan can also protect cilok from bacteria and fungi. oxidation and bacterial contamination. On Day 4 there was a significant Cilok fish at the third treatment, change. The colour of the fish cilok cooked cilok then drained and stored at changed from white to brownish. On Day room temperature. Cilok fish is not given a 5, there were changes in the texture of the chitosan coating or mixture. Based on the fish cilok, from chewy to inelastic. Such three types of treatment, later it can be changes happened because the fish cilok observed which cilok is more durable in started being oxidated. storage, so that it can be seen which chitosan treatment is more suitable to be D1 D2 used as cilok natural preservative.

Test Result Data 1. Cilok with 1,5% chitosan coating Data from the observation towards fish D3 D4 D5 cilok with 1,5% chitosan coating is presented in Table 3 and Figure 3 below.

Table 3. The Result Data of the Observation on Fish Cilok with 1,5% Chitosan Coating Day Parameters Observation Figure 3. The Condition of Cilok with 1,5% results Chitosan Coating. (1) (2) (3) D1 Taste Fish flavoured 2. The cilok mixed with 1,5% chitosan Colour White solution Odor/aroma Fish flavoured The result data of the observation on Texture Chewy Taste Fish flavoured the fish cilok mixed with a 15 cc of 1,5% D2 Colour White chitosan solution is presented in Table 4 Odor/aroma Fish flavoured and Figure 4. Texture Chewy Based on the results of the Taste Fish flavoured observations on Day 1 and Day 2, there D3 Colour White Odor/aroma Fish flavoured were no significant changes in taste, Texture Chewy colour, odor and texture. On Day 3, change Taste Fish flavoured occured in the texture, from chewy to D4 Colour Brownish inelastic. It was caused by the fact that the Odor/aroma Fish flavoured fish cilok started getting oxidated. On Day Texture Chewy Taste Fish flavoured 4, the colour changed from white to D5 Colour Brownish brown, the odor went bad and the texture Odor/aroma Fish flavoured became inelastic. On Day 5, the cilok Texture Inelastic tasted sour and slightly bitter, had brownish colour, smelt bad and became Based on the observation on Day 1, inelastic. These changes were caused by there were no significant change in taste, the fact that the fish cilok had been colour, odor and texture since fungi and oxidated and bacteria and fungi started bacteria had not infected the fish cilok. The growing in them. same situation were observed on Day 2 and 3 since the fish cilok with 1,5% chitosan coating in cold temperature for two minutes were able to delay oxidation

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Table 4. The Result Data of The Observation on mushy. These changes were results of the Fish Cilok mixed with 1,5% chitosan oxidation and the growth of bacteria and solution fungi in the cilok. Day Parameters Observation results On Day 4, the odor got worse, the taste (1) (2) (3) became sour and slightly bitter, the colour Taste Fish flavoured turned to brownish and the texture became D1 Colour White slimy and mushy. These were caused by Odor/aroma Fish flavoured the increasing contaminations by fungi and Texture Chewy Taste Fish flavoured bacteria. On Day 5, the cilok tasted bitter, D2 Colour White had blackish and brownish colour, smelt Odor/aroma Fish flavoured bad, and had mushy texture due to the Texture Chewy growth of fungi and bacteria. This shows Taste Fish flavoured that the cilok without chitosan spoiled fast. D3 Colour White Odor/aroma Fish flavoured Texture Inelastic Table 5. The Result Data of The Observation on Taste Fish flavoured the Fish Cilok without Chitosan D4 Colour Brownish Day Parameters Observation Odor/aroma Started rotting results Texture Inelastic (1) (2) (3) Taste Sour to slightly Taste Fish flavoured D5 bitter D1 Colour White Colour Brown Odor/aroma Fish flavoured Odor/aroma Rotten Texture Chewy Texture Very inelastic Taste Fish flavoured D2 Colour Brownish Odor/aroma Fish flavoured D1 D2 Texture Mushy Taste Sour to slightly D3 bitter Colour Brownish Odor/aroma Started rotting Texture Mushy D3 D4 D5 Taste Sour to slightly D4 bitter Colour Brownish Odor/aroma Rotten Texture Mushy Taste Bitter Figure 4. The Condition of Cilok Mixed with a 15 D5 Colour Blackish brown cc of 1,5% chitosan solution. Odor/aroma Rotten Texture Mushy 3. The fish cilok without chitosan The result data of the observation on the fish cilok without chitosan is presented D1 D2 in Table 5 and Figure 5. Based on the observation result on Day 1, the cilok had normal fishy taste, white colour, fishy odor and chewy texture. On Day 2, changes happened. The colour D3 D4 D5 changed from white to brownish and the texture changed from chewy to slightly mushy. On Day 3, changes occured in the taste. They tasted sour and slightly bitter, the odour smell bad, and the texture stayed Figure 5. The Condition of Cilok without Chitosan

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CONCLUSION Membrane of Gram-Negative Bacteria. International Journal of Food Microbiology, 71: 235–244. The conclusions of this research are as follows: (1) Chitosan can be produced by Killay, A. (2013). Kitosan Sebagai Anti processing shrimp shells and heads Bakteri pada Bahan Pangan yang Aman dan Tidak Berbahaya. through demineralisation, deproteinization Prosiding FMIPA Universitas and deacetylation. (2) Fish cilok can be Pattimura 2013 – ISBN: 978-602- made from Treadfin Bream (Nemipterus 97522-0-5. Nematophorus). (30 Chitosan is a natural Marganov. (2003). Potensi Limbah Udang preservative for fish cilok. It is proven by sebagai Penyerap Logam Berat the results of the experiment and (Timbal, Kadmium, dan Tembaga) observations that the fish cilok with 1,5% di Perairan. (Disertasi). Program chitosan coating were able to last longer Pascasarjana IPB, Bogor. up to fourth day preservation (D+3) in a Safira, M. (2014). Cilok, Jajanan Gurih room temperature. Kenyal yang Rawan Dicampur Bahan Aditif. Retrieved from REFERENCES http://food.detik.com/read/2014/06/ 25/175123/2619297/1345/cilok- Agustina, S., Swantara, I.M.D., & Suartha, jajanan-gurih-kenyal-yang-rawan- I.N. (2015). Isolasi Kitin, dicampur-bahan-aditif Karakterisasi, dan Sintesis Kitosan dari Kulit Udang. Jurnal Kimia 9 Sanjaya, I., & Yuanita, L. (2007). (2), Juli 2015: 271-278. Adsorpsi Pb(II)oleh Kitosan Hasil Isolasi Kitin CangkangKepiting Arifin, Zainal & Nugroho, P. (2016). Bakau (Scylla). Jurnal Ilmu Dasar, Limbah Udang Sebagai Pengawet 8(1): 30-36. Ikan Patin (Pangasius sp.). Prosiding Seminar Nasional Teknik Silvia, R., Wahyu W, S., Hanum, S. Kimia “Kejuangan” ISSN 1693- (2014). Pemanfaatan Kitosan dari 4393, 17 Maret 2016. Cangkang Rajungan (Portonus Sanginolentus L.) Sebagai Pengawet Deviana, A. (2015). Boraks. Retrieved Ikan Kembung (Rastrelliger Sp) dan from Ikan Lele (Clarias Batrachus). http://www.academia.edu/31109269 Jurnal Teknik Kimia USU, Vol. 3, /BORAKS No. 4 Desember 2014.

Dompeipen, E.J., Kaimudin, M., & Dewa, Soegiarto, R.A., Purwijantiningsih, R.P. (2016). Isolation Chitin and L.M.E., & Pranata, S. (2013). Chitosan From Waste of Skin Aplikasi Kitosan Sebagai Pengawet Shrimp. Majalah BIAM 12 (01) Alami dari Kulit Udang Dogol (2016) 32-38. (Metapenaeus monoceros Fab.) pada Sosis Daging Sapi. (Skripsi). Hafdani, F.N. & Sadeghinia. N., (2011). A Program Studi Biologi Universitas Review on Application of Chitosan Atma Jaya Yogyakarta. as a Natural Antimicrobial. International Journal of Suprapti,L. (2003). Membuat: Pharmacological and Daging & Bakso Ikan. Yogyakarta: Pharmaceutical Sciences Vol:5, Kanisius. No:2, 2011 Suptijah, P., Salamah, E., Sumaryanto, H., Harjanti, R.S. (2014). Kitosan dari Limbah Purwaningsih, S., & Santoso, J. Udang Sebagai Bahan Pengawet (1992). Pengaruh Berbagai Metode . Jurnal Rekayasa Isolasi Khitin Udang Terhadap Proses, Vol. 8 No. 1, 2014. Mutunya. Laporan Penelitian Jurusan Pengolahan Hasil Perikanan Helander, I.M., Nurmiaho-Lassila, E.L., Fakultas Perikanan IPB, Bogor. Ahvenainen, R., Rhoades J., & Roller, S. (2001). Chitosan Disrupts Trisnawati, E., Dewid, A., & Saleh, A. The Barrier Properties of The Outer (2013). Pembuatan Kitosan dari

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Limbah Cangkang Kepiting Sebagai Yanuarlan, J. (2015). Gurih Kenyal Cilok, Bahan Pengawet Buah Duku dengan Aci Dicolok. Retrieved from Variasi Lama Pengawetan. Jurnal http://www.wewengkonsumedang.c Teknik Kimia No. 2, Vol. 19, April om/2015/01/gurih-kenyal-cilok-aci- 201. dicolok.html.

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