Ekstraksi, karakterisasi dan aktivitas antioksidan hyaluronan dari vitreous humor mata tuna sirip kuning (Thunnus albacares)

Wini Trilaksani, Bambang Riyanto, Tri Wahyuningsih


Vitreous humor is a part of the tuna fish eye that plays a role in maintaining the brightness of the eye and contains hyalocytes having function to produce hyaluronan. Hyaluronan is thought as an antioxidant to prevent unsaturated fatty acids damage in tuna fish eye caused by free radicals. Hyaluronan from vitreous humor tuna fish eye has not been widely studied. This study was aimed to extract the hyaluronan found in vitreous humor of tuna fish eye and determine its characteristics. Extraction of hyaluronan consisted of presipitation, centrifugation, enzyme digestion, defatting, dialysis, and freeze drying. Antioxidant activity was determined by three methods comprising 1,1-diphenyl-2-picrylhydrazyl2,2-azinobis (DPPH), 3-ethylbenzo-thiozoline-6-sulfonic acid (ABTS), and ferric reducing antioxidant power (FRAP). Vitreous humor tuna fish eye had a proportion of 31% of eye, pH value 6.905±0.0083, viscosity 195.56±12.86 cP, water 98.09±0.49%, and glutamic acid 932.06±3.80 mg/L. The hyaluronan yield was about 0.005±0.001%. The IC50 antioxidant activity of hyaluronan against DPPH was 25.52±0.22 mg/mL while the inhibition capacity on ABTS was 1.92±0.08% and the ferric reduction capacity was 9.34±0.55 µM/g. These results indicates that the antioxidant activity of hyaluronan from vitreous humor tuna fish eye is low.


Amagai I, Tashiro Y, Ogawa H. 2009. Improvement of the extraction procedure for hyaluronan from fish eyeball and the molecular characterization. Fisheries Science. 75(3): 805-810.
[AOAC] Association of Official Analytical and Chemistry. 1999. Officials Methods of Analysis 18th Edition. Maryland (US): AOAC International.
[ASTM] American Society for Testing Material. 2010. ASTM D789: Standard Test Methods for Determination of Solution Viscosities of Polyamide (PA). Pennsylvania (US) : American Society for Testing Material.
[ASTM] American Society for Testing Material. 2013. ASTM E1252: Standard Practice for General Techniques for Obtaining Infrared Spectra for Qualitative Analysis. Pennsylvania (US): American Society for Testing Material.
Benzie IFF, Strain JJ. 1996. The ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: the FRAP assay. Analytical Biochemistry. 239(1): 70-76.
Bishop P. 1996. The biochemical structure of mammalian vitreous. Eye. 10: 664-670.
Blois MS. 2005. Antioxidant determination by the use of stable free radical. Nature. 181:1191- 1200.
Brand-Williams W, Cuvelier ME, Berset C. 1995. Use of a free radical method to evaluate antioxidant activity. Lebensmittel-Wissenschaft und-Technologie. 28(1): 25-30.
[BSN] Badan Standardisasi Nasional. 1992. Cara Uji Makanan dan Minuman Standar Nasional Indonesia : SNI 01-2891-1992. Jakarta (ID): Badan Standardisasi Nasional.
Budria RJ, Gonzalez EF, Sanz HMT, Ostkiz SJL.1989.Amniotic fluid phospholipids : study of the cold acetone precipitation effect. Clinica Chimica Acta.180 (2): 141-146.
Campo GM, Avenoso A, Campo S, D’Ascola A, Ferlazzo AM, Calatroni A. 2004. The antioxidant and antifibrogenic effects of the glycosaminoglycans hyaluronic acid and chondroitin-4-sulphate in a subchronic rat model of carbon tetrachloride-induced liver fibrogenesis. Chemico-Biological Interactions. 148(3): 125-138.
Carneiro, Döll-Boscardin PM, Fiorin BC, Nadal JM, Farago PV, de Paula JP. 2016. Development and characterization of hyaluronic acid-lysine nanoparticles with potential as innovative dermal filling. Brazilian Journal of Pharmaceutical Sciences. 52(4): 645-651.
Choi J, Kim JK, Kim JH, Kweon DK, Lee JW. 2010. Degradation of hyaluronic acid powder by electron beam irradiation, gamma ray irradiation, microwave irradiation and thermal treatment: A comparative study. Carbohydrate Polymers. 79: 1080-1085.
Cowman MK, Lee H G , Schwertfeger KL , McCarthy JB Turley EA. 2015. The content and size of hyaluronan in biological fluids and tissues. Frontiers in Immunology. 6:261. 1-8.
Donati S, Caprani SM, Airaghi G, Vinciguerra R, Bartalena L, Testa F, Mariotti C, Porta G, Simonelli F, Azzolini C. 2014. Vitreous substitutes: the present and the future. BioMed Research International. 2014: 1-12.  
Duke-elder WS. 1929. The Physico-chemical properties of the vitreous body. The Journal of Physiology. 68 (2): 155-165.
Falco F, Barra M, Cammarata M, Cuttitta A, Jia S, Bonanno A, Mazzola S, Wu G. 2016. Amino acid composition in eyes from zebrafish (Danio rerio) and sardine (Sardina pilchardus) at the larval stage. SpringerPlus. 26:5:519.
Fatt I, Weissman BA. 1992. Physiology of The Eye : An Introduction to The Vegetative Functions. Los Angeles (US) : Reed Publishing Inc.
Farmer JG, Benomran F, Watson AA, Harland WA. 1985. Magnesium, potassium, sodium and calcium in post-mortem vitreous humour from humans. Forensic Science International. 27(1): l-13.
Flemming CR. 1989. Trace element metabolism in adult patients requiring total parenteral nutrition. The Aflemmmerican Journal of Clinical Nutrition. 49 (3): 573–579.
Gamarro EG, Orawattanamateekul W, Sentina J, dan Gopal TKS. 2013. By products of tuna processing. Rome (IT): Globefish Research Programme.
Garg HG, Hales CA. 2004. Chemistry and biology of hyaluronan. London (UK) : Elsevier.
[Globefish.org] Global Fish Economy. 2017. Global market trends for raw and processed tuna remained uncertain throughout 2016 (diakses pada 20 April).http://www.fao.org/in-action/globefish/market-reports/resource-detail/en/c/882595/.
Hafsa J, Chaouch MA, Charfeddine B, Rihouey C, Limem K, Cerf DL, Rouatbi S, Majdoub H. 2017. Effect of ultrasonic degradation of hyaluronic acid extracted from rooster comb on antioxidant and antiglycation activities. Pharmaceutical Biology. 55 (1): 156-163.
Haxaire K, Maréchal Y, Milas M, Rinaudo M. 2003. Hydration of polysaccharide hyaluronan observed by IR spectrometry. I. Preliminary experiments and band assignments. Biopolymers. 72 (1): 10-20.
Horkay F, Basser PJ, Londono DJ, Hech AM, Geissler E. 2009. Ions in hyaluronic acid solutions. The Journal of Chemical Physics. 131 (18): 1-8.
Kanchana K, Arumugam M, Giji S, Balasubramanian T. 2013. Isolation, characterization and antioxidant activity of hyaluronic acid from marine bivalve mollusc Amussium pleuronectus (Linnaeus, 1758). Bioactive Carbohydratesand Dietary Fibre. 2 (1): 1 – 7.
Khoddami A, Ariffin AA, Bakar J, Ghazali HM. 2012. Quality and fatty acid profile of the oil extracted from fish waste (head, intestine and liver) (Euthynnus affinis).African Journal of Biotechnology. 11(7): 1683-1689.
[KKP] Kementerian Kelautan dan Perikanan. 2018. Pesona tuna sebagai penggerak bisnis perikanan Indonesia. (diakses pada 28 September 2018) http://kkp.go.id/artikel/4409-pesona- tuna-sebagai-penggerak-bisnis-perikanan-Indonesia.
Kim JK, Srinivasan P, Kim JH, Choi J, Park HJ, Byun MW, Lee JW. 2008. Structural and antioxidant properties of gamma irradiated hyaluronic acid. Food Chemistry. 109 (4): 763-770.
Lapcik L, Omelka L, Kubena K, Galatik A, Kello V. 1990. Photodegradation of hyaluronic acid and of the vitreous body. General Physiology and Biophysics. 9 (4): 419-429.
Lapuerta P, Schein SJ. 1995. A four-surface schematic eye of macaque monkey obtained by an optical method. Vision Research. 35 (16): 2245-2254.
Li S, Xiong Q, Lai X, Li X, Wan M, Zhang J, Yan Y, Cao M, Lu L, Guan J, Zhang D, Lin Y. 2016. Molecular modification of polysaccharides and resulting bioactivities. Comprehensive Reviews Food Sciencean and Food Safety. 15: 237–250.
Lorget F, Parenteau A, Carrier M, Lambert D, Gueorguieva A, Schuetz C, Bantseev V, Thackaberry E. 2015. Characterization of the pH and temperature in the rabbit, pig, and monkey eye: key parameters for the development of long-acting delivery ocular strategies. Molecular Pharmacetics. 13 (9):1-6.
Meyer K, Palmer JW. 1934. The polysaccharide of the vitreous humor. Journal Biology Chemistry.107: 629-634.
Merce ALR, Carrera LCM, Romanholi LKS, Recio MAL. 2002. Aqueous and solid complexes of iron(III) with hyaluronic acid : Potentiometric titrations and infrared spectroscopy studies. Journal of Inorganic Biochemistry. 89 (3-4): 212-218. 
Morgano MA, Rabonato LC, Milani RF, Miyagusku L, Balian SC. 2011. Assessment of trace elements in fishes of Japanese foodsmarketed in Sa˜o Paulo (Brazil). Food Control. 22: 778–785.
Morris ER, Rees DA, Welsh EJ. 1980. Conformation and dynamic interactions in hyaluronate solutions. Journal Molecular Biology. 138 (2): 383–400.
Moseley R, Walker M, Waddington RJ, Chen WYJ. 2003. Comparison of the antioxidant properties of wound dressing materials–carboxymethylcellulose, hyaluronan benzyl ester and hyaluronan, towards polymorphonuclear leukocyte-derived reactive oxygen species. Biomaterials. 24 (9): 1549-1557.
Murado MA, Montemayor MI, Cabo ML, Vasquez JA, Gonzalez MP. 2012. Optimization of extraction and purification process of hyaluronic acid from fish eyeball. Food and Bioproducts Processing. 90 (3): 491-498.
Mutamimah D, Ibrahim B, Trilaksani W. 2018. Antioxidant activity of protein hydrolysate produced from tuna eye (Thunnus sp.) by enzymatic hydrolysis. Jurnal Pengolahan Hasil Perikanan Indonesia. 21(3): 522-531.
Mutti D, Zadnik K, Murphy CJ. 1999. Naturally occurring vitreous chamber—based myopia in the labrador retriever. Investigative Ophthalmology & Visual Science. 40(7):1577-84.
Necas J, Bartosikova L, Brauner P, Kolar J. 2008. Hyaluronic acid (hyaluronan): a review. Veterinarni Medicina. 53 (8): 397-411.
Nollet LML. 1996. Handbook of Food Analysis Amino Acid. New York (US): Marcel DekkerInc.
Peng S, Chen C, Shi Z, Wang L. 2013. Amino acid and fatty acid composition of the muscle tissue of yellowfin tuna (Thunnus albacares) and bigeye tuna (Thunnus obesus). Journal of Food and Nutrition Research. 1(4): 42-45.
Prasertsan P, Prachumratana T. 2008. Comparison and selection of protease and lipase sources from visceral organs of three tuna species. Songklanakarin Journal of Science and Technology. 30(1) : 73-76.
Prior RL, Wu X, Schaich K. 2005. Standardized methods for determination of antioxidant capacity and phenolics in foods and dietary supplements. Journal Agricultural and Food Chemistry. 53(10): 4290-4302.
Polson C, Sarkar P, Incledon B, Raguvaran V, Grant R. 2003. Optimization of protein precipitation based upon effectiveness ofprotein removal and ionization effect in liquid chromatography–tandem mass spectrometry. Journal of Chromatography B. 785 (2) : 263-275.
Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Evans CR. 1999. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine. 26 (9-10): 1231-1237.
Reddy KJ, Karunakaran KT. 2013. Purification and characterization of hyaluronic acid produced by Streptococcus zooepidemicus strain 3523-7. Journal of Bioscience and Biotechnology. 2(3): 173-179.
Renuka V, Anandan R, Suseela M, Ravishankar CN, Sivaraman GK. 2016. Fatty acid profile of yellowfin tuna eye (Thunnus albacares) and oil sardine muscle (Sardinella longiceps). Fishery Technology. 53: 151 – 154.
Rinjani. 2017. Potensi pemanfaatan limbah industri perikanan tuna. [Skripsi]. Bogor (ID): Institut Pertanian Bogor.
Schneider A, Picart C, Senger B, Schaaf P, Voegel JC, Frisch B. 2007. Layer-by-layer films from hyaluronan and amine modified hyaluronan. National Instituties of Health Public Access. 23(5): 2655-2662.
Scott JE. 1992. The chemical morphology of the vitreous. Eye. 6 : 553-555.
Sebag J. 1989. The Vitreous : Structure, Function, and Pathobiology. California (US) : Springer-Verlag New York Inc.
Sebag J. 1998. Macromolecular structure of corpus vitreus. Progress in Polymer Science. 23 (3): 415 - 446.
Soltês L, Mendhici R, Kogan G, Schiller J, Stankovska M, Arnhold J. 2006. Degradative action of reactive oxigen species on hyaluronan. Biomacromolecules. 7(3): 659-668.
Soman N, Banerjee R. 2003. Artificial vitreous replacements. Biomedical Materials and Engineering. 13 (1): 59-74.
Suri S, Banerjee R. 2006. Biophysical evaluation of vitreous humor, its constituents and substitutes. Trends Biomaterial Artifisial Organs. 20(1): 72-77.
Toppe J. 2017. Fish for food security – more than just fillets. INFOFISH International. 4: 18-20.
Ürgeová E, Vulganová K. 2016. Comparison of enzymatic hydrolysisof polysaccharides from eggshells membranes. Nova Biotechnologica et Chimica.15 (2): 133-140.
Vasi AM, Popa MI, Butnaru M, Dodi G, Verestiuc L. 2014. Chemical functionalization of hyaluronic acid for drug deliveryapplications. Materials Science and Engineering C. 38: 177-185.
Verlag GmbH, Weinheim. 2003. Handbook of Spectroscopy. Jerman (DE): Wiley-VCH.
Volpi N, Maccari F. 2003. Purification and characterization of hyaluronic acid from the mollusc bivalve Mytilus galloprovinciali. Biochimie. 85 (6): 619–625.
Young RG, Williams HH. 1954. Biochemistry of the eye : gelatinous protein of vitreous body. Archives of Ophthalmol. 51(5):593-595.


Wini Trilaksani
wtrilaksani@gmail.com (Primary Contact)
Bambang Riyanto
Tri Wahyuningsih
TrilaksaniW., RiyantoB., & Wahyuningsih T. (2019). Ekstraksi, karakterisasi dan aktivitas antioksidan hyaluronan dari vitreous humor mata tuna sirip kuning (Thunnus albacares). Jurnal Pengolahan Hasil Perikanan Indonesia, 22(3). https://doi.org/10.17844/jphpi.v22i3.29126

Article Details