Abstract
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.
Highlights
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
Extraction of hyaluronan consisted of presipitation, centrifugation, enzyme digestion, defatting, dialysis, and freeze drying
Summary
Bahan utama yang digunakan dalam penelitian berupa mata ikan tuna beku jenis tuna Thunnus albacares yang diperoleh dari industri pengolahan ikan tuna Muara Baru, Jakarta Utara, yang berasal dari penangkapan perairan Samudera Hindia pada bulan Januari sampai April tahun 2017, kemudian disimpan beku suhu -18oC. Pelarut yang digunakan untuk ekstraksi adalah setilpiridinium klorida (CPC) (Merck) 3%, natrium klorida (NaCl) 0,4 M, kalium asetat 10%, etanol pro analis, buffer asetat 0,1 M (pH 4,5 dan 3,6), asam trikloroasetat (TCA) 10%, akuades, aseton, enzim papain komersil (Merck), dan kantung dialisis ukuran 5 nm (5 KDa) (Sigma aldrich). Bahan pengujian antioksidan berupa asam askorbat 98,09%, metanol pro analis, 1,1-difenil-2-picrylhydrazyl (DPPH Merck), 2,2-azinobis Sulfonic acid) (ABTS Sigma Aldrich), kalium persulfat 99%, Besi (III) klorida (FeCl3). (Merck), 2,4,6-tripiridil-s-triazin (TPTZ), asam klorida (HCl), dan FeSO4 (Merck). Alat yang digunakan berupa timbangan analitik (Satorius BS-24S), kertas saring nilon 100 mesh, pH meter
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