Characterization, Antioxidant Activity, and In Silico Molecular Docking of Chitosan from Snail Shell Waste by Ultrasonic Technique

Umarudin1,2 Sri Rahayu3, Sri Widyarti3, Warsito4,5
1 Doctoral student of Biology, Faculty of Mathematic and Natural Sciences, University of Brawijaya, Malang 65145, East Java, Indonesia. 2 Department of Pharmacy, Diploma III Pharmacy, Academy Pharmacy of Surabaya, Surabaya 60231, East Java, Indonesia. 3 Department of Biology, Faculty of Mathematics and Natural Sciences, University of Brawijaya, Malang 65145, East Java, Indonesia. 4 Department of Chemistry, Faculty of Mathematics and Natural Sciences, University of Brawijaya, Malang 65145, East Java, Indonesia. 5 Essential Oil's Institute, University of Brawijaya, Malang 65145, East Java, Indonesia.
PDF icon full_text.pdf818.04 KB
Snails are often found in Indonesia, especially at Kediri, but the snail shell has no commercial value. This research report describes the characterization and antioxidant activity of chitosan from snail shell waste (chitosan-SSW) by ultrasonic technique and analyzes the potential of chitosan as an inhibitor of receptors of free radicals using an in silico molecular docking method. Characterization of chitosan-SSW was performed to analyze the content of water, protein, and functional groups as well as molecular weight, particle size, mor-phology, antioxidant activity, and in silico molecular docking. We found that chitosan-SSW with ultrasonic treatment had a high degree of deacetylation (DD) and high molecular weight (MW). The characteristic of chitosan-SSW was found to be as follows: water content of 0.43 %, protein content of 1.59 %, molecular weight of 2.198 kDa, and deacetylation degree value of 79.50 %. Importantly, chitosan-SSW had high antioxidant activity to potentially reduce free radical of DPPH with IC50 value of 2.44 µg/mL. Chitosan is predicted to have the potential as an inhibitor of lipoxygenase, CYP2C9, and NADPH-oxidase.

[1] Bedoić, R.; Ćosić, B.; Duić, N. Technical Potential and

Geographic Distribution of Agricultural Residues, Co-Products and By-Products in the European Union. Sci. Total Environ. 2018, 686, 568-579.

[2] Vanitha, C.; Kuppusamy, M.R.; Sridhar, T.M.; Sureshkumar, R.; Mahalakshmi, N. Synthesis Characterization of Nano-Hydroxy Apatite From White Snail Shells and Removal of Methylene Blue. Int. J. Innov. Res. Adv. Eng. 2017, 4, 2014-2018.

[3] Oyekunle, D.T; Omoleye, J. A. Effect of Particle Sizes on the Kinetics of Demineralization of Snail Shell for Chitin Synthesis Using Acetic Acid. Heliyon 2019, 5, 1-7.

[4] Oyekunle, D.T; Omoleye, J. Extraction, Characterization and Kinetics of Demineralised of Chitin Produced From Snail Shells of Different Particle Sizes Using 1.2M HCL. Int. J. Mech. Eng.

Technol. 2019, 10, 2010-2020.

[5] Xu, R.; Mao, J.; Penh, N.; Luo, X.; Chang, C. Chitin/clay

Microspheres with Hierarchical Architecture for Highly Efficient Removal of Organic Dyes. Carbohydr. Polym. 2018, 188, 143-150.

[6] Popadyuk, N.; Zholobko, O.; Donchak, V.; Harhay, K.;

Budishevska, O.; Voronov, A.; Kohut, A.; Voronov, S. Ionically and Covalently Crosslinked Hydrogel Particles Based on Chitosan and Poly (ethylene glycol). Chem. Chem. Technol. 2014, 8,


[7] Bazunova, M.; Sharafutdinova, L.; Bazunova, A.; Lazdin, R.; Elinson, M.; Kulish, E. Biocompatible Gel-like Forms of Drugs on the Basis of Solutions of Polysaccharide Chitosan with Alcohols. Chem. Chem. Technol. 2018, 12, 43-46.

[8] Neha, K.; Anitha, R.; Subashini, R.; Natarajan, A.; Sridha. T. M. Synthesis and Characterization of Chitosan/Potato Peel Powder-Based Hydrogel and its in vitro Antimicrobial Activity. J. Appl. Pharm. Sci. 2019, 9, 66-71.

[9] Solomko, N.; Budishevska, O.; Voronov, S. Peroxide Chitosan Derivatives and their Application. Chem. Chem. Technol. 2007, 1, 137-147.

[10] Umarudin; Rahayu, S.; Warsito.; Widyarti, S. Molecular

Characterization, Antioxidant, And Toxicity Activity Of Chitosan Isolated From Lissahatina Fulica Shell Waste Using Hot Plate

Magnetic Stirrer Technique. Rasayan J. Chem. 2022, 15, 2299-2303.

[11] Umarudin; Widyarti, S.; Warsito; Rahayu, S. Effect of

Lissachatina Fulica Chitosan on the Antioxidant and Lipid Profile of Hypercholesterolemic Male Wistar Rats. J. Pharm. Pharmacogn. Res. 2022, 10, 995-1005.

[11] Sharma, K.; Somavarapu, S.; Colombani, A.; Govind, N.; Taylor, K. M.G. Crosslinked Chitosan Nanoparticle Formulations for Delivery from Pressurized Metered Dose Inhalers. Eur. J. Pharm. Biopharm. 2012, 81, 74-81.

[12] Carocho, M; Ferreira, I.C.F.R. A review on Antioxidants, Prooxidants and Related Controversy: Natural and Synthetic

Compounds, Screening and Analysis Methodologies and Future Perspectives. Food Chem. Toxicol. 2013, 51, 15-25.

[13] Kancheva, V. D. Phenolic Antioxidants-Radical-Scavenging and Chain-Breaking Activity: A Comparative Study. Eur. J. Lipid Sci. Technol. 2009, 111, 1072-1089.

[14] Yuliana, A.; Pradeckta, L.S.; Savitri, E.; Handaratri, A.R.; Sumarno. The Effect of Sonication on the Characteristic of

Chitosan. Proceeding of International Conference on Chemical and Material Engineering 2012, 1-5.

[15] Albu, S.; Joyce, E; Paniwnyk, L; Lorimer, J.P.; Mason, T.J. Potential for the Use of Ultrasound in the Extraction of Antioxidants from Rosmarinus Officinalis for the Food and Pharmaceutical Industry. Ultrason. Sonochem. 2004, 11, 261-265.

[16] Zhang, Q.-W.; Lin, L.-G.; Ye, W.-C. Techniques for Extraction and Isolation of Natural Products: A Comprehensive Review. Chinese Med. 2018, 13, 20.

[17] AOAC. Official Methods of Analysis, 18th edn. Washington, DC: Association of Official Analytical Chemists, 2007.

[18] Xuan Du, D.; Xuan Vuong, B. Study on Preparation of

Water-Soluble Chitosan with Varying Molecular Weights and Its Antioxidant Activity. Adv. Mater. Sci. Eng. 2019, 2019, 8781013.

[19] Journot, C.M.A.; Nicolle, L.; Lavanchy, Y.; Gerber-Lamaire, S. Selection of Water-Soluble Chitosan by Microwave-Assisted

Degradation and pH-Controlled Precipitation. Polymers 2020, 12, 1274.

[20] Hsu, C.-Y.; Chan, Y.-P.; Chang, J. Antioxidant Activity of Extract from Polygonum cuspidatum. Biol. Res. 2007, 40, 12-21. https://doi:10.4067/S0716-97602007000100002

[21] Wafiroh, S.; Wathoniyyah, M.; Abdulloh, A.; Rahardjo, Y.; Fahmi, M. A. Application of Glutaraldehyde-Crosslinked Chitosan Membranes from Shrimp Shellwaste on Production of Biodiesel from Calophyllum Inophyllum Oil. Chem. Chem. Technol. 2017, 11, 65-70.

[22] Oyekunle, D. T; Omoleye, J. A. E. New Process for

Synthesizing Chitosan from Snail Shells. J. Phys. Conf. Ser. 2019, 1299, 012089.

[23] [EFSA] European Food Safety Authority. Scientific Opinion on the Safety of Chitinglucan as a Novel Food Ingredient. EFSA J. 2011, 9, 2137.

[24] Ningrum, S.R.; Sinaga, S.M.; Harahap, U. Isolation of Chitosan from Cuttlefish Bones. Int. J. Sci. Technol. Manag. 2022, 3,


[25] Yuan, Y.; Wan, Z.-L.; Yin, S.-W.; Teng, Z.; Yang, X.-Q.; Qi, J.-R.; Wang, X.-Y. Formation and Dynamic Interfacial Adsorption-of Glycinin/Chitosan Soluble Complex at Acidic pH:

Relationship to Mixed Emulsion Stability. Food Hydrocoll. 2013, 31, 85-93.

[26] Kusumaningsih, T.; Masykur, A.; Arief, U. Synthesis of

hitosan from the Chitin Of Escargot (Achatina fulica). Biofarmasi Journal of Natural Product Biochemistry 2004, 2, 64-68.

[27] Waryani, S.W.; Silvia, R.; Hanum, F. Utilization of Chitosan from The Shells of Snail (Achatina fulica) as a Preservative of Plush Fish (Rastrelliger sp) and Catfish (Clarias batrachus). Jurnal Teknik Kimia 2014, 3, 51-57.

[28] Hossain, M.S; Iqbal, A. Production and Characterization of Chitosan from Shrimp Waste. J. Bangladesh Agric. Univ. 2014, 12, 153-160.

[29] Srinivasan, H.; Kanayairam, V.; Ravichandran, R. Chitin and Chitosan Preparation from Shrimp Shells Penaeus Monodon and its Human Ovarian Cancer Cell Line, PA-1. Int. J. Biol. Macromol. 2018, 107, 662-667.

[30] Xuan Du, D.; Xuan Vuong, B. Study on Preparation of

Water-Soluble Chitosan with Varying Molecular Weights and Its Antioxidant Activity. Adv. Mater. Sci. Eng. 2018, 1-8.

[31] Zhang, H.; Li, Y.; Zhang, X; Liu, B.; Zhao, H.; Chen, D. Directly Determining the Molecular Weight of Chitosan with Atomic Force Microscopy. Front Nanosci. Nanotech. 2016, 185, 57-63.

[32] Lu, C.; Li, H.; Li, C.; Chen, B.; Shen, Y. Chemical

Composition and Radical Scavenging Activity of Amygdalus

pedunculata Pall Leaves Essential Oil. Food Chem. Toxicol. 2018, 19, 368-374.

[33] Ma, Y.-L.; Zhu, D.-Y.; Thakur, K.; Wang, C.-H.; Wang, H.; Ren, Y.-F.; Zhang, J.-G.; Wei, Z.-J. Antioxidant and Antibacterial Evaluation of Polysaccharides Sequentially Extracted from Onion (Allium cepa L.). Int. J. Biol. Macromol. 2018, 111, 92-101.

[34] Prakakash, P.; Neelu, G. Therapeutic Uses of Ocimum Santum Linn (Tulsi) with a Note on Eugenol and its Pharmacological Ac-tions: A Short Review. Indian J. Physiol. Pharmacol. 2005, 49, 125-131.

[35] Jafari, H.; Bernaerts, K. V.; Dodi. G.; Shavandi. A.

Chitooligosaccharides for Wound Healing Biomaterials Engineer-ing. Mater. Sci. Eng. C 2020, 117, 111266. https://doi:10.1016/j.msec.2020.111266

[36] Ngo, D.-H.; Kim, S.-K. Chapter Two - Antioxidant Effects Of Chitin, Chitosan, and their Derivatives. Adv. Food Nutr. Res. 2014, 73, 15-31. https://doi:10.1016/b978-0-12-800268-1.00002-0

[37] Rahayu, S.; Prasetyawan, S.; Suprihatin, T.; Ciptadi, G. In-silico study of Marselia crenata compounds as activator Keap1/Nrf2 pathway in ovarian function. IOP Conf. Ser.: Earth Environ. Sci. 2021, 743, 012056.