Antioxidant Activity of Hydrolysate Protein in Skipjack Fish (Katsuwonus pelamis) Using the ABTS Method
DOI:
https://doi.org/10.22487/j24775185.2026.v15.i2.pp142-149Keywords:
ABTS, antioxidants, bromelain, protein hydrolysate, skipjack fish bonesAbstract
Skipjack tuna (Katsuwonus pelamis) bones are a byproduct of the fishing industry that are rich in protein and have potential as a source of bioactive peptides. This study aimed to determine the antioxidant activity of protein hydrolysates from raw and cooked skipjack tuna bones using the ABTS method. Hydrolysis was performed using bromelain at concentrations of 4%, 6%, and 8% at 55 °C for 4 hours pH 7.0), followed by enzyme inactivation at 85 °C for 20 minutes. Antioxidant activity was measured by IC50 determination using a UV-Vis spectrometer at 734 nm. The results showed that the antioxidant activity of skipjack tuna bone samples with the highest total protein content was observed at an enzyme concentration of 8%, namely 35.55% (raw fish bones) and 36.24% (cooked fish bones). The highest degree of hydrolysis in the raw fish bone sample reached 82.15 % and in the cooked fish bone sample reached 86.81 % with 8 % enzyme. Qualitative amino acid tests (ninhydrin, Biuret, xanthoprotein, and Millon) yielded positive results, indicating the presence of aromatic amino acids (tyrosine, tryptophan, phenylalanine) that contribute to antioxidant activity. The lowest IC50 value was obtained in the raw fish bone sample at an 8 % concentration, namely 10.03 mg/ml, and in the cooked fish bone sample at 6.19 mg/ml at an 8 % enzyme concentration. The higher the enzyme concentration, the higher the antioxidant activity.
References
Andhikawati, A., & Pratiwi, D. Y. (2021). A review: Methods of smoking for the quality of smoked fish. Asian Journal of Fisheries and Aquatic Research, 13(4), 37–43. https://doi.org/10.9734/ajfar/2021/v13i430273
AOAC. (2005). Official methods of analysis. Maryland: AOAC International.
Bastı´as, J. M., Balladares, P., Acuña, S., Quevedo, R., & Muñoz, O. (2017). Determining the effect of different cooking methods on the nutritional composition of salmon (salmo salar) and chilean jack mackerel (trachurus murphyi) fillets. Plos One, 12(July), 1–10. https://doi.org/10.1371/journal.pone.0180993
Botutihe, D. N., Hudiyono, S., & Saepudin, E. (2024). In vitro antioxidant and hypocholesterolemic potency of melon (cucumis melo l.) seed protein hydrolysate. Current Research in Nutrition and Food Science, 12(2), 792-804. http://dx.doi.org/10.12944/CRNFSJ.12.2.24
BPS. (2024). Produksi Tanaman Buah-buahan, 2021–2023. Jakarta: Badan Pusat Statistik.
Dwiningrum, R., Pisacha, I. M., & Nursoleha, E. (2019). Review : Analisis kualitatif dan kuantitatif kandungan protein pada olahan bahan pangan. Jurnal Farmasi, 2(2), 60–67. https://doi.org/doi.org/10.33378/jppik.v13i2.195
Farhan, M., Hasani, I. W., Khafaga, D. S. R., Ragab, W. M., Kazi, R. N. A., Aatif, M., Muteeb, G., & Fahim, Y. A. (2025). Enzymes as catalysts in industrial biocatalysis: Advances in engineering, applications, and sustainable integration. Catalysts, 15(9), 1–31. https://doi.org/10.3390/catal15090891
Hadji, F., Botutihe, D. N., Najmah, N., Bialangi, N., & Kunusa, W. R. (2025). Aktivitas antioksidan hidrolisat protein tulang ikan selar bentong (selar crumenophthalmus) hasil hidrolisis enzim papain. Jurnal Crystal : Publikasi Penelitian Kimia dan Terapannya, 7(2), 277–285. https://doi.org/10.36526/jc.v7i2.6186
Hermaya, A. A., Edison, E., & Diharmi, A. (2021). Aktivitas antioksidan hidrolisat protein ikan cunang (congresox talabon). Jurnal Agroindustri Halal, 7(1), 79–86. https://doi.org/10.30997/jah.v7i1.3593
Idowu, A. T., Benjakul, S., Sinthusamran, S., Sookchoo, P., & Kishimura, H. (2019). Protein hydrolysate from salmon frames: Production, characteristics and antioxidative activity. Journal of Food Biochemistry, 43(2), 1–12. https://doi.org/10.1111/jfbc.12734
Ilyas, N. M., Setiasih, S., & Hudiyono, S. (2020). Isolasi dan karakterisasi enzim bromelain dari bonggol dan daging buah nanas (ananas comosus). Jurnal Ilmiah Kimia dan Pendidikan Kimia, 21(2), 133–141. https://doi.org/10.35580/chemica.v21i2.17983
Jafar, I., Asfar, M., & Mahendradatta, M. (2025). Characterization and antioxidant activity of protein hydrolysate from milkfish (chanos chanos). Food Science and Preservation, 32(6), 1042–1053. https://doi.org/10.11002/fsp.2025.32.6.1042
Korczek, K., Tkaczewska, J., & Migdał, W. (2018). Antioxidant and antihypertensive protein hydrolysates in fish products - a review. Czech Journal of Food Sciences, 36(3), 195–207. https://doi.org/10.17221/283/2017-CJFS
Meilani, S. S., Kustiyah, E., Saing, B., & Ridwan, A. M. (2022). Pemanfaatan kembali limbah tulang ikan tenggiri sebagai bahan baku pembuatan gelatin melalui proses hidrolisis asam fosfat. Jurnal Lemuru: Ilmu Perikanan dan Kelautan, 4(2), 57–64. https://doi.org/10.36526/lemuru.v4i2.2085
Mutamimah, D., Ibrahim, B., & Trilaksani, W. (2018). Antioxidant activity of protein hydrolysate produced from tuna eye (thunnus sp.) by enzymatic hydrolysate. Jurnal Pengolahan Hasil Perikanan Indonesia, 21(3), 522–531. https://doi.org/10.17844/jphpi.v21i3.24736
Noman, A., Wang, Y., Zhang, C., & Abed, S. M. (2022). Antioxidant activity of hybrid sturgeon (huso dauricus × acipenser schrenckii) protein hydrolysate prepared using bromelain, its fractions and purified peptides. Polish Journal of Food and Nutrition Sciences, 72(1), 79–89. https://doi.org/10.31883/pjfns/146317
Nugraha, A. A. (2022). Hidrolisis protein kacang kedelai dengan enzim bromelain dan aktivitas antikanker terhadap sel kanker payudara MCF-7. Unpublished undergraduate’s thesis. Jakarta: UIN Syarif Hidayatullah.
Prastyo, D. T., Trilaksani, W., & Nurjanah. (2020). Aktivitas antioksidan hidrolisat kolagen kulit ikan nila (oreochromis niloticus). Jurnal Pengelolaan Hasil Perikanan Indonesia, 23(3), 423–433. https://doi.org/10.17844/jphpi.v23i3.31732
Pratama, A. N., & Busman, H. (2020). Potensi antioksidan kedelai (glycine max l) terhadap penangkapan radikal bebas. Jurnal Ilmiah Kesehatan Sandi Husada, 11(1), 497–504. https://doi.org/10.35816/jiskh.v11i1.333
Puspawati, N. M., Dewi, P. P., Bogoriani, N. W., & Ariati, N. K. (2020). Produksi hidrolisat protein antioksidan melalui hidrolisis enzimatik protein kulit ayam broiler dengan enzim papain. Jurnal Kimia, 14(2), 206-212. https://doi.org/10.24843/JCHEM.2020.v14.i02.p16
Rahmawati, H., Agustini, T. W., Dewi, E. N., & Trianto, A. (2024). Karakteristik hidrolisat kolagen kulit ikan tenggiri kering dengan enzim papain. Jurnal Pengolahan Hasil Perikanan Indonesia, 27(12), 1156–1171. https://doi.org/10.17844/jphpi.v27i12.55831
Ratnayani, K., Listiyanti, N. K. L., Ariati, N. K., & Laksmiwati, A. A. I. A. M. (2024). Monitoring hidrolisis protein kecambah kacang tunggak (vigna unguiculata l.) oleh enzim alkalase pada variasi waktu dan rasio enzim-substrat. Jurnal Kimia, 18(2), 145-152. https://doi.org/10.24843/JCHEM.2024.v18.i02.p07
Sari, K., Rahmadi, A., Rohmah, M., Saragih, B., & Salam, I. (2024). Antioxidant activities (DPPH and ABTS method) from extract of Bangle rhizome (zingiber cassumunar) using different method of extraction. AcTion: Aceh Nutrition Journal, 9(1), 110–119. http://dx.doi.org/10.30867/action.v9i1.1531
Sbroggio, M. F., Montilha, M. S., Figueiredo, V. R. G. D., Georgetti, S. R., & Kurozawa, L. E. (2016). Influence of the degree of hydrolysis and type of enzyme on antioxidant activity of okara protein hydrolysates. Food Science and Technology, 36(2), 375–381. https://doi.org/10.1590/1678-457X.000216
Sholahuddin, M. A., Dyah, N., Lastuti, R., & Amin, M. (2024). Effect of differences beomelain enzyme concentration on protein hydrolysat from waste of tilapia viscera (oreochromis sp.) on antioxidant activity. Jurnal Biosains Pascasarjana, 26(1), 15–22. https://doi.org/10.20473/jbp.v26i1.2024.15-22
Sholahuddin, M. A., & Prayoga, A. (2023). The development of antioxidant products from Tilapia offal protein hydrolysate with different enzyme concentrations and a review in fulfillment of the halal product assurance criteria. JHPR: Journal of Halal Product and Research, 6(2), 138–146. https://doi.org/10.20473/jhpr.vol.6-issue.2.138-146
Susanty, A., & Kusumaningrum, I. (2021). Pengaruh waktu hidrolisis terhadap karakteristik hidrolisat protein ikan toman (channa micropeltes) asal DAS Kalimantan Timur. Jurnal Riset Teknologi Industri, 15(2), 463–475. https://doi.org/10.26578/jrti.v15i2.7462
Syahfitri, A., Daulay, A. S., Ridwanto, & Yuniarti, R. (2025). Analysis of protein content of some types of milk high in protein using the kjeldahl and visible spectrophotometry method. Journal of Pharmaceutical and Sciences, 8(2), 847–858. https://doi.org/10.36490/journal-jps.com.v8i2.865
Taniyo, W., Salimi, Y. K., & Iyabu, H. (2021). Karekteristik dan aktivitas antioksidan hidrolisat protein ikan nike (awaous melanocephalus). Jurnal Pendidikan Kimia dan Ilmu Kimia, 4(2), 52-63. https://dx.doi.org/10.31602/dl.v4i2.5935
Tumanggor, J. S. B., Razoki, Yunus, M., & Harahap, D. W. S. (2025). Uji Aktivitas Antioksidan Ekstrak Etanol Herba Sirih Cina ( Peperomia pellucida L .) dengan Metode DPPH. Jurnal Ilmu Kedokteran dan Kesehatan Indonesia, 5(2), 241–248. https://doi.org/10.55606/jikki.v5i2.6142
Varilla, C., Marcone, M., Paiva, L., & Baptista, J. (2021). Bromelain, a group of pineapple proteolytic complex enzymes (ananas comosus) and their possible therapeutic and clinical effects. a summary. Foods, 10(10), 1–14. https://doi.org/10.3390/foods10102249
Warda, S. A., Botutihe, D. N., Arviani, A., Musa, W. J. A., & Iyabu, H. (2025). Aktivitas antioksidan hidrolisat protein kepala dan kulit udang vename ( litopenaeus vannamei). Jurnal Crystal: Publikasi Penelitian Kimia dan Terapannya, 7(2), 267–276. https://doi.org/10.36526/jc.v7i2.6189
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Fou F. Muksin, Deasy N. Botutihe, Erni Mohamad, Hendri Iyabu, Najmah Najmah
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.
