• NABANITA GHOSH School of Community Science and Technology (SOCSAT), Indian Institute of Engineering Science and Technology, Shibpur, Howrah, India.
  • BHATTACHARYYA DK School of Community Science and Technology (SOCSAT), Indian Institute of Engineering Science and Technology, Shibpur, Howrah, India.


Objective: Isolation, characterization and analysis of antioxidant activity of protein extracted from Labeo rohita skin.

Methods: The present work aimed to characterize protein isolated from Indian major carp L. rohita skin. Protein isolation was carried out by the salting-out method and the protein content was estimated with bovine serum albumin. Isolated protein was subjected to Fourier-transform infrared analyzer to identify the presence of –NH3 group to be claimed as protein. It was also subjected to high-performance liquid chromatography (HPLC) analyzer and run against standard collagen (Sigma), the molecular weight of the protein was determined through mass spectroscopy (MS) to know the protein structure more precisely the protein isolate was studied under a scanning electron microscope (SEM). Antioxidative activity of the crude protein sample was studied through 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant potential (FRAP) assay, 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay.

Results: The protein isolated was 23% of the dry weight of fish skin. In HPLC analysis, the isolated protein gives peaks that are similar to that of standard collagen, and in MS, its molecular weight was near about 300 kDa, which is the molecular weight of collagen type 1. The SEM image shows a crystal structure of protein isolates. Results of antioxidative property show that fish skin protein isolates have good antioxidative activity also.

Conclusion: The isolated protein was collagen and it can be used as a replacer of the renowned market available collagen.

Keywords: Protein, Collagen, Fish skin, 2,2-Diphenyl-1-picrylhydrazyl, ABTS, FRAP


1. Zhang F, Wang A, Li Z, He S, Shao L. Preparation and characterization of collagen from freshwater fish scales. Food Nut Sci 2011;2:818-23.
2. Sudharsan S, Seedevi P, Saravanan R, Ramasamy P, Kumar SV, Vairamani S, et al. Isolation, characterization and molecular weight determination of collagen from marine sponge Spirastrella inoconstans (Dendy). Arf J Biotech 2013;12:504-11.
3. Blanco M, Vázquez JA, Pérez-Matín RI, Sotelo CG. Extraction optimization from the skin of the small-spotted catshark (S.canicula) by response surface methodology. Mar Drugs 2019;17:1-14.
4. Morimura S, Uemura Y, Kida K, Nagata H, Fahmi A, Shigematsu T. Development of an effective process for utilization of collagen from livestock and fish waste. Process Biochem 2002;37:1403-12.
5. Kolodziejska I, Sikorski ZE, Knwska N. Parameters affecting the isolation of collagen from squid (Illex argentimes) skins. Food Chem 1999;66:153-7.
6. Nagai T, Suzuki N. Preparation and characterization of several fish bone collagen. J Food Biochem 2000;24:427-36.
7. Swatscheck D, Schattoon W, Kellermann J, Mullar WE, Kreuter J. Marine sponge collagen: Isolation, characterization and effects on the skin parameter surface-ph, moisture and sebum. Eur J Pharm Biopharm 2002;53:107-13.
8. Sadowska M, Kolodziejska I, Niecikowska C. Isolation of collagen from the skins of Baltic cod (Gadus morhua). Food Chem 2003;81: 257-62.
9. Vijayan DK, Sreerekha PR, Tejpal CS, Asha KK, Mathew S, Ravishankar CN, et al. Extraction and characterization of acid soluble collagen (ASC) from air bladder of striped cat fish (Pangasius hypophthalmus). Int J Fish Aquat Stud 2018;6:310-8.
10. Ghosh N, Bhattacharyya DK. Effect of heating time and heat on the physicochemical and antioxidative property of fish (Labeo rohita) skin oil. Int J Pharm Pharm Sci 2019;11:87-9.
11. Ghosh N, Roy M, Ghosh M, Bhattacharyya DK. Study on Physicochemical and Antioxidant Properties of Blend of Fish Skin (Labeo rohita) Oil and Chia Seed (Salvia hispanica) Oil. Kolkata: 3rd International conference on Biotechnology and Biological Sciences Biospectrum; 2019. p. 121-6.
12. Riaz T, Zeeshan R, Zarif F, Ilyas K, Muhammad N, Safi SZ, et al. FT-IR analysis of natural and synthetic collagen. Appl Spectro Rev 2018;53:703-46.
13. Belbachir K, Noreen R, Gouspillou G, Petibois C. Collagen types analysis and differentiation by FTIR spectroscopy. Anal Bioanal Chem 2009;395:829-37.
14. Pataridis S, Eckhardt A, Mikulíková K, Sedláková P, Miksík I. Determination and quantification of collagen types in tissues using HPLC-MS/MS. Curr Analyt Chem 2009;5 316-23.
15. Noitup P, Garnjanagoonchorn W, Morrissey, MT. Fish skin Type I collagen. J Aquat Food Prod Technol 2005;14:17-28.
16. Sujithra S, Kiruthiga N, Prabhu MJ, Kumeresan R. Isolation and determination of Type I collagen from tilapia (Oreochromis niloticus) waste. Int J Eng Technol 2013;5:2181-5.
99 Views | 136 Downloads
How to Cite
GHOSH, N., and B. DK. “ISOLATION, CHARACTERIZATION, AND DETERMINATION OF ANTIOXIDATIVE PROPERTIES OF PROTEIN EXTRACTED FROM ROHU (LABEO ROHITA) FISH SKIN”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 13, no. 3, Mar. 2020, pp. 79-83, doi:10.22159/ajpcr.2020.v13i3.36615.
Original Article(s)