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Production of Water-Soluble Chitosan from Crab Shells (Portunus sp.) by Pressurized Hydrolysis Method as an Active Material for Hand Sanitizer
Corresponding Author(s) : Niken Dhamayanti
Jurnal Ilmiah Perikanan dan Kelautan, 2024: IN PRESS ISSUE (JUST ACCEPTED MANUSCRIPT, 2024)
Abstract
Graphical Abstract
Highlight Research
- Water-soluble chitosan from crab’s shell can be produced via the pressurized hydrolysis method in an acidic environment with a pressure cooker; the optimal treatment is a 3% HCl concentration.
- Water-soluble chitosan from crab’s shell has the following properties: it is yellowish white in color, odorless, powder-like, with a yield of 83.37±0.73, an acidity degree of 5, 83±0.34, viscosity of 69.0±0.82, solubility of 93.57±0.33, and a degree of deacetylation of 78.4%.
- The optimal concentration for water-soluble chitosan from crab’s shell inhibition is 160 mg/ml, with an inhibition zone of 7.47 mm for S. aureus and 6.70 mm for E. coli, falling in the medium category.
- Physical features of organoleptic water-soluble chitosan from crab’s shell hand sanitizer: neutral appearance (5), somewhat similar fragrance (6), neutral texture (5), similar (6), not homogenous, dispersion 3.59-4.03 cm, pH 6.05-6.28.
- The most effective hand sanitizer formulation from crab’s shell water soluble chitosan is HS3 (including 200 mg/ml of water-soluble chitosan), which has a weak inhibition zone of 5.35±0.57 mm for S. aureus bacteria and 4.70±0.07 mm for E. coli.
Abstract
The study explores the production of water-soluble chitosan (WSC) from crab shells (Portunus spp.) through a pressurized hydrolysis method. The limited solubility of chitosan at neutral pH restricts its usability. Natural WSC can serve as an active antibacterial component in hand sanitizers. Therefore, the aim of this study was to produce WSC from crab shells using pressurized hydrolysis methods as an active ingredient for hand sanitizer. Chitosan was depolymerized into WSC by utilizing hydrochloric acid (2, 3, and 4%) and was hydrolyzed using a pressure cooker at a temperature of approximately 110˚C for 1 hour. Isopropyl alcohol was then added to the filtrate at a ratio of 2:1. The selected water-soluble chitosan was treated with 3% HCl and made into 3 different concentrations of 140, 150, and 160 mg/ml, then tested for its antibacterial activity. The WSC hand sanitizer antibacterial test has concentrations of 180, 190 and 200 mg/ml, and for positive control using commercial hand sanitizer, and negative control in the form of basic gel without chitosan. By depolymerizing chitosan using 3% HCl, a high solubility (93.57±0.33) of WSC was achieved, with a degree of deacetylation (DD) value of 78.4%. The results indicated that the concentration of water-soluble chitosan is160 mg/ml and exhibited the strongest inhibition against S. aureus and E. coli, with clear area values of 7.47 mm and 6.70 mm, respectively. The best hand sanitizer formulation is HS3 (addition of water-soluble chitosan 200 mg/ml) and the ability to inhibit S. aureus bacteria with a clear area value of 5.35 ± 0.57 mm and E. coli is 4.70 ± 0.07 mm. Further research may explore the scalability of pressure hydrolysis, and the broad-spectrum antibacterial efficacy of chitosan produced from crab shells.
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- Achmad, H., Djais, A. I., Jannah, M., Carmelita, A. B., Uinarni, H., Arifin, E. M., & Putra, A. P. (2020). Antibacterial Chitosan of Milkfish Scales (Chanos chanos) on Bacteria Prophyromonas gingivalis & Agregatibacter actinomycetemcomitans. Systematic Reviews in Pharmacy, 11(6): 836-841.
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- Aranaz I, Alcántara AR, Civera MC, Arias C, Elorza B, Heras Caballero A, Acosta N. Chitosan: An Overview of Its Properties and Applications. Polymers (Basel). 2021 Sep 24;13(19):3256.
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- Buijs, N. P., Matheson, E. J., Cochrane, S. A., & Martin, N. I. (2023). Targeting Membrane-bound Bacterial Cell Wall Precursors: A Tried-and-True Antibiotic Strategy in Nature and the Clinic. Chemical Communications, 59(50):7685-7703.
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- Chamidah, A., Widiyanti, C. H., & Fabiyani, N. N. (2016). Utilization of Water-Soluble Chitosan as an Antiseptic Hand Sanitizer. Jurnal Perikanan Universitas Gadjah Mada, 21(1):9-16.
- Das, A., Ringu, T., Ghosh, S., & Pramanik, N. (2023). A Comprehensive Review on Recent Advances in Preparation, Physicochemical Characterization, and Bioengineering Applications of Biopolymers. Polymer Bulletin, 80(7):7247-7312.
- Dwiyitno, D., Basmal, J., & Mulyasari, M. (2017). Effect of Esterification Temperature on the Characteristics of Carboxymethyl Chitosan (Cmcts). Jurnal Penelitian Perikanan Indonesia, 10(3):67-73.
- Egorov, A. R., Kirichuk, A. A., Rubanik, V. V., Rubanik Jr, V. V., Tskhovrebov, A. G., & Kritchenkov, A. S. (2023). Chitosan and its Derivatives: Preparation and Antibacterial Properties. Materials, 16(18):1-29.
- Faustine, D., Setyaningsih, I., & Hardiningtyas, S. D. (2020). Chitosan Depolymerization Using Ultraviolet Light and Hydrochloric Acid Catalyst. Jurnal Pengolahan Hasil Perikanan Indonesia, 23(3):412-422.
- Feng, Z., Adolfsson, K. H., Xu, Y., Fang, H., Hakkarainen, M., & Wu, M. (2021). Carbon Dot/polymer nanocomposites: From Green Synthesis to Energy, Environmental and Biomedical Applications. Sustainable Materials and Technologies, 29(206):1-25.
- Ghimire, U., Kandel, R., Ko, S. W., Adhikari, J. R., Kim, C. S., & Park, C. H. (2024). Electrochemical Technique to Develop Surface-Controlled Polyaniline Nano-Tulips (PANINTs) on PCL-reinforced Chitosan Functionalized (CS-f-Fe2O3) Scaffolds for Stimulating Osteoporotic Bone Regeneration. International Journal of Biological Macromolecules, 264(48):1-18.
- Gumilar, J., Suryaningsih, L., & Setia, D. F. (2023). The Use of Various Hydrochloric Acid Concentration Levels on Rabbit Bone Gelatin Quality. Jurnal Ilmu Ternak Universitas Padjadjaran, 23(2):154-160.
- Hahn, T., Tafi, E., Paul, A., Salvia, R., Falabella, P., & Zibek, S. (2020). Current State of Chitin Purification and Chitosan Production from Insects. Journal of Chemical Technology & Biotechnology, 95(11):2775-2795.
- Hayati, R., Sari, A., Hanum, F., Nabilah, N., Earlia, N., & Lukitaningsih, E. (2023). Formulation and Antibacterial Activity of Averrhoa bilimbi L. Fruits Extract in Vegetable Oil-based Liquid Hand Soap. Malacca Pharmaceutics, 1(1):30-36.
- Huang, W. (2019). Multifunctional Self-healing Hydrogels Based on Natural Polymers for Biomedical Applications. [Thesis]. University of Alberta. Canada.
- Jin, T., Liu, T., Lam, E., & Moores, A. (2021). Chitin and Chitosan on the Nanoscale. Nanoscale Horizons, 6(7):505-542.
- Kadak, A. E., & Salem, M. O. A. (2020). Antibacterial Activity of Chitosan, Some Plant Seed Extracts and Oils Against Escherichia Coli and Staphylococcus Aureus. Alınteri Journal of Agriculture Sciences, 35(2):144-150.
- Kadak, A. E., Küçükgülmez, A., & Çelik, M. (2023). Preparation and Characterization of Crayfish (Astacus leptodactylus) Chitosan with Different Deacetylation Degrees. Iranian Journal of Biotechnology, 21(2):87-94.
- Ke CL, Deng FS, Chuang CY, Lin CH. Antimicrobial Actions and Applications of Chitosan. Polymers (Basel). 2021 Mar 15;13(6):904.
- Kusrini, E., Wilson, L. D., Padmosoedarso, K. M., Mawarni, D. P., Sufyan, M., & Usman, A. (2023). Synthesis of Chitosan Capped Zinc Sulphide Nanoparticle Composites as an Antibacterial Agent for Liquid Handwash Disinfectant Applications. Journal of Composites Science, 7(2):1-17.
- Li, N., Xiong, X., Ha, X., & Wei, X. (2019). Comparative Preservation Effect of Water-soluble and Insoluble Chitosan from Tenebrio Molitor Waste. International Journal of Biological Macromolecules, 133 (15 July 2019):165-171.
- Li, Q., Dunn, E. T., Grandmaison, E. W., & Goosen, M. F. (2020). Applications and Properties of Chitosan. In Applications of Chitin and Chitosan (pp. 3-29). CRC Press.
- Manus N, Yamlean P.V.Y, Kojong N. S. (2016) Formulation of Citronella Leaf Essential Oil Gel (Cymbopogon citratus) as Hand Antiseptic. Pharmaceutical Scientific Journal (Pharmacon), 5 (3):85-93.
- Meata, B. A., Uju, U., & Trilaksani, W. (2019). Characteristics of Glucosamine Hydrochloride Result of Chitosan Hydrolysis Using Acid and Ultrasonication. Jurnal Pascapanen dan Bioteknologi Kelautan dan Perikanan, 14(2):151-162.
- Mohan K, Ganesan AR, Ezhilarasi PN, Kondamareddy KK, Rajan DK, Sathishkumar P, Rajarajeswaran J, Conterno L. Green and eco-friendly approaches for the extraction of chitin and chitosan: A review. Carbohydr Polym. 2022 Jul 1; 287:119349.
- Oktarlina, R. Z., Bahri, S., & Adjeng, A. N. T. (2022). Production and Characterization of Micro-collagen from Carp Scales Waste (Cyprinus carpio). Research Journal of Pharmacy and Technology, 15(5):1995-2002.
- Natalia, D. A., Dharmayanti, N., & Dewi, F. R. (2021). Chitosan Production from Crab Shells (Portunus Sp.) at Room Temperature. Jurnal Pengolahan Hasil Perikanan Indonesia, 24(3):301-309.
- Pambudi, G. B. R., Ulfin, I., Harmami, H., Suprapto, S., Kurniawan, F., & Ni'Mah, Y. L. (2018). Synthesis of water-soluble chitosan from crab shells (Scylla serrata) waste. In F. Kurniawan, Y. Kusumawati, S. Fatmawati, H. Juwono, & A. S. Purnomo (Eds.), 3rd International Seminar on Chemistry: Green Chemistry and Its Role for Sustainability Article 020086. AIP Conference Proceedings; 2049(2018):1551-7616.
- Pellis, A., Guebitz, G. M., & Nyanhongo, G. S. (2022). Chitosan: Sources, Processing and Modification Techniques. Gels, 8(7):1-27.
- Permadi, A., Afifah, R. A., Apriani, D. A. K., & Ariyani, F. (2022). Water Soluble Chitosan from Green Mussel (Perna viridis) Shells and its Use as Fat-absorber in Cookies. Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology, 17(3):168-176.
- Peter, S., Lyczko, N., Gopakumar, D., Maria, H. J., Nzihou, A., & Thomas, S. (2021). Chitin and Chitosan Based Composites for Energy and Environmental Applications: A Review. Waste and Biomass Valorization, 12(2021):4777-4804.
- Robiatun, R. R., Pangondian, A., Paramitha, R., Zulmai Rani, & Gultom, E. D. (2022). Formulation And Evaluation Of Hand Sanitizer Gel From Clove Flower Extract (Eugenia aromatica L.). International Journal of Science, Technology & Management, 3(2):484-491. https://doi.org/10.46729/ijstm.v3i2.472
- Saheed, I. O., Da Oh, W., & Suah, F. B. M. (2021). Chitosan Modifications for Adsorption of Pollutants–a Review. Journal of Hazardous Materials, 408(15):1-61.
- Saputra, D., Jamilah, J., & Lahay, N. (2023). Antibacterial Inhibition Test of Butterfly Pea (Clitoria ternatea L.) as Alternative Feed Additive for Broiler. AIP Conference Proceedings.
- Somwanshi, S. B., Kumar, S., & Rathore, G. S. (2023). Cosmetic Science. (1st ed) AG Publishing House (AGPH Books). 241 p.
- Sudianto., Suseno, S. H., & Suptijah, P. (2020). Optimization of Water-Soluble Chitosan Production Using the Pressure Hydrolysis Method. Jurnal Pengolahan Hasil Perikanan Indonesia, 23(3):441-446.
- Wang, W., Xue, C., & Mao, X. (2020). Chitosan: Structural Modification, Biological Activity and Application. International Journal of Biological Macromolecules, 164(2020):4532-4546.
- Wang, J., & Zhuang, S. (2022). Chitosan-based Materials: Preparation, Modification and Application. Journal of Cleaner Production, 355(2022):1- 25.
- Wulandari, I. O., Pebriatin, B. E., Valiana, V., Hadisaputra, S., Ananto, A. D., & Sabarudin, A. (2022). Green Synthesis of Silver Nanoparticles Coated by Water Soluble Chitosan and its Potency as Non-alcoholic Hand Sanitizer Formulation. Materials, 15(13):1-20.
- Zhou, J., Cai, Y., Liu, Y., An, H., Deng, K., Ashraf, M. A., Zou, L., & Wang, J. (2022). Breaking Down the Cell Wall: Still an Attractive Antibacterial Strategy. Frontiers in Microbiology, 13(2022):1-21.
References
Achmad, H., Djais, A. I., Jannah, M., Carmelita, A. B., Uinarni, H., Arifin, E. M., & Putra, A. P. (2020). Antibacterial Chitosan of Milkfish Scales (Chanos chanos) on Bacteria Prophyromonas gingivalis & Agregatibacter actinomycetemcomitans. Systematic Reviews in Pharmacy, 11(6): 836-841.
Ahmad, A., & Ayub, H. (2022). Fourier Transform Infrared Spectroscopy (FTIR) Technique for Food Analysis and Authentication. In Nondestructive quality assessment techniques for fresh fruits and vegetables Singapore: Springer Nature 11(03):1582-1856.
Aranaz I, Alcántara AR, Civera MC, Arias C, Elorza B, Heras Caballero A, Acosta N. Chitosan: An Overview of Its Properties and Applications. Polymers (Basel). 2021 Sep 24;13(19):3256.
Ardean, C., Davidescu, C. M., Nemeş, N. S., Negrea, A., Ciopec, M., Duteanu, N., Negrea, P., Duda-seiman, D., & Muntean, D. (2021). Antimicrobial Activities of Chitosan Derivatives. Pharmaceutics, 13(10):12-21.
Ardean, C., Davidescu, C. M., Nemeş, N. S., Negrea, A., Ciopec, M., Duteanu, N Negrea, P., Duda-seiman, D., & Musta, V. (2021). Factors Influencing the Antibacterial Activity of Chitosan and Chitosan Modified by Functionalization. International Journal of Molecular Sciences, 22(14):12-28.
Arifin, S. H. A. G. (2021). Formulation, Physical Stability Test, And Antimicrobial Activity Of Gel Hand Sanitizer From Combination of Piper Betle and Moringa Oleifera Leaves Extract. Universitas Islam Negeri Sunan Ampel. Surabaya.
Ariyanthini, K. S., Angelina, E., Permana, K. N. B., Thelmalina, F. J., & Prasetia, I. G. N. J. A. (2021). Antibacterial Activity Testing of Hand Sanitizer Gel Extract of Coriander (Coriandrum sativum L.) Seeds Against Staphylococcus aureus. Journal of Pharmaceutical Science and Application, 3(2):98-107.
Azmana, M., Mahmood, S., Hilles, A. R., Rahman, A., Arifin, M. A. B., & Ahmed, S. (2021). A Review on Chitosan and Chitosan-based Bionanocomposites: Promising Material for Combatting Global issues and its applications. International journal of biological macromolecules, 185(31):832-848.
Bahri, S., Ginting, Z., Vanesa, S., & Nasrul, Z. A. (2021). Formulation of Patchouli Plant Essential Oil Gel (Pogostemon Cablin Benth) as a Hand Antiseptic (Hand Sanitizer). Jurnal Teknologi Kimia Unimal, 10(1):87-99.
Buijs, N. P., Matheson, E. J., Cochrane, S. A., & Martin, N. I. (2023). Targeting Membrane-bound Bacterial Cell Wall Precursors: A Tried-and-True Antibiotic Strategy in Nature and the Clinic. Chemical Communications, 59(50):7685-7703.
Cahyono, E. (2018). Characteristics of Chitosan from Tiger Prawn (Panaeus monodon) Shell Waste. Akuatika Indonesia, 3(2):96-102.
Chamidah, A., Widiyanti, C. H., & Fabiyani, N. N. (2016). Utilization of Water-Soluble Chitosan as an Antiseptic Hand Sanitizer. Jurnal Perikanan Universitas Gadjah Mada, 21(1):9-16.
Das, A., Ringu, T., Ghosh, S., & Pramanik, N. (2023). A Comprehensive Review on Recent Advances in Preparation, Physicochemical Characterization, and Bioengineering Applications of Biopolymers. Polymer Bulletin, 80(7):7247-7312.
Dwiyitno, D., Basmal, J., & Mulyasari, M. (2017). Effect of Esterification Temperature on the Characteristics of Carboxymethyl Chitosan (Cmcts). Jurnal Penelitian Perikanan Indonesia, 10(3):67-73.
Egorov, A. R., Kirichuk, A. A., Rubanik, V. V., Rubanik Jr, V. V., Tskhovrebov, A. G., & Kritchenkov, A. S. (2023). Chitosan and its Derivatives: Preparation and Antibacterial Properties. Materials, 16(18):1-29.
Faustine, D., Setyaningsih, I., & Hardiningtyas, S. D. (2020). Chitosan Depolymerization Using Ultraviolet Light and Hydrochloric Acid Catalyst. Jurnal Pengolahan Hasil Perikanan Indonesia, 23(3):412-422.
Feng, Z., Adolfsson, K. H., Xu, Y., Fang, H., Hakkarainen, M., & Wu, M. (2021). Carbon Dot/polymer nanocomposites: From Green Synthesis to Energy, Environmental and Biomedical Applications. Sustainable Materials and Technologies, 29(206):1-25.
Ghimire, U., Kandel, R., Ko, S. W., Adhikari, J. R., Kim, C. S., & Park, C. H. (2024). Electrochemical Technique to Develop Surface-Controlled Polyaniline Nano-Tulips (PANINTs) on PCL-reinforced Chitosan Functionalized (CS-f-Fe2O3) Scaffolds for Stimulating Osteoporotic Bone Regeneration. International Journal of Biological Macromolecules, 264(48):1-18.
Gumilar, J., Suryaningsih, L., & Setia, D. F. (2023). The Use of Various Hydrochloric Acid Concentration Levels on Rabbit Bone Gelatin Quality. Jurnal Ilmu Ternak Universitas Padjadjaran, 23(2):154-160.
Hahn, T., Tafi, E., Paul, A., Salvia, R., Falabella, P., & Zibek, S. (2020). Current State of Chitin Purification and Chitosan Production from Insects. Journal of Chemical Technology & Biotechnology, 95(11):2775-2795.
Hayati, R., Sari, A., Hanum, F., Nabilah, N., Earlia, N., & Lukitaningsih, E. (2023). Formulation and Antibacterial Activity of Averrhoa bilimbi L. Fruits Extract in Vegetable Oil-based Liquid Hand Soap. Malacca Pharmaceutics, 1(1):30-36.
Huang, W. (2019). Multifunctional Self-healing Hydrogels Based on Natural Polymers for Biomedical Applications. [Thesis]. University of Alberta. Canada.
Jin, T., Liu, T., Lam, E., & Moores, A. (2021). Chitin and Chitosan on the Nanoscale. Nanoscale Horizons, 6(7):505-542.
Kadak, A. E., & Salem, M. O. A. (2020). Antibacterial Activity of Chitosan, Some Plant Seed Extracts and Oils Against Escherichia Coli and Staphylococcus Aureus. Alınteri Journal of Agriculture Sciences, 35(2):144-150.
Kadak, A. E., Küçükgülmez, A., & Çelik, M. (2023). Preparation and Characterization of Crayfish (Astacus leptodactylus) Chitosan with Different Deacetylation Degrees. Iranian Journal of Biotechnology, 21(2):87-94.
Ke CL, Deng FS, Chuang CY, Lin CH. Antimicrobial Actions and Applications of Chitosan. Polymers (Basel). 2021 Mar 15;13(6):904.
Kusrini, E., Wilson, L. D., Padmosoedarso, K. M., Mawarni, D. P., Sufyan, M., & Usman, A. (2023). Synthesis of Chitosan Capped Zinc Sulphide Nanoparticle Composites as an Antibacterial Agent for Liquid Handwash Disinfectant Applications. Journal of Composites Science, 7(2):1-17.
Li, N., Xiong, X., Ha, X., & Wei, X. (2019). Comparative Preservation Effect of Water-soluble and Insoluble Chitosan from Tenebrio Molitor Waste. International Journal of Biological Macromolecules, 133 (15 July 2019):165-171.
Li, Q., Dunn, E. T., Grandmaison, E. W., & Goosen, M. F. (2020). Applications and Properties of Chitosan. In Applications of Chitin and Chitosan (pp. 3-29). CRC Press.
Manus N, Yamlean P.V.Y, Kojong N. S. (2016) Formulation of Citronella Leaf Essential Oil Gel (Cymbopogon citratus) as Hand Antiseptic. Pharmaceutical Scientific Journal (Pharmacon), 5 (3):85-93.
Meata, B. A., Uju, U., & Trilaksani, W. (2019). Characteristics of Glucosamine Hydrochloride Result of Chitosan Hydrolysis Using Acid and Ultrasonication. Jurnal Pascapanen dan Bioteknologi Kelautan dan Perikanan, 14(2):151-162.
Mohan K, Ganesan AR, Ezhilarasi PN, Kondamareddy KK, Rajan DK, Sathishkumar P, Rajarajeswaran J, Conterno L. Green and eco-friendly approaches for the extraction of chitin and chitosan: A review. Carbohydr Polym. 2022 Jul 1; 287:119349.
Oktarlina, R. Z., Bahri, S., & Adjeng, A. N. T. (2022). Production and Characterization of Micro-collagen from Carp Scales Waste (Cyprinus carpio). Research Journal of Pharmacy and Technology, 15(5):1995-2002.
Natalia, D. A., Dharmayanti, N., & Dewi, F. R. (2021). Chitosan Production from Crab Shells (Portunus Sp.) at Room Temperature. Jurnal Pengolahan Hasil Perikanan Indonesia, 24(3):301-309.
Pambudi, G. B. R., Ulfin, I., Harmami, H., Suprapto, S., Kurniawan, F., & Ni'Mah, Y. L. (2018). Synthesis of water-soluble chitosan from crab shells (Scylla serrata) waste. In F. Kurniawan, Y. Kusumawati, S. Fatmawati, H. Juwono, & A. S. Purnomo (Eds.), 3rd International Seminar on Chemistry: Green Chemistry and Its Role for Sustainability Article 020086. AIP Conference Proceedings; 2049(2018):1551-7616.
Pellis, A., Guebitz, G. M., & Nyanhongo, G. S. (2022). Chitosan: Sources, Processing and Modification Techniques. Gels, 8(7):1-27.
Permadi, A., Afifah, R. A., Apriani, D. A. K., & Ariyani, F. (2022). Water Soluble Chitosan from Green Mussel (Perna viridis) Shells and its Use as Fat-absorber in Cookies. Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology, 17(3):168-176.
Peter, S., Lyczko, N., Gopakumar, D., Maria, H. J., Nzihou, A., & Thomas, S. (2021). Chitin and Chitosan Based Composites for Energy and Environmental Applications: A Review. Waste and Biomass Valorization, 12(2021):4777-4804.
Robiatun, R. R., Pangondian, A., Paramitha, R., Zulmai Rani, & Gultom, E. D. (2022). Formulation And Evaluation Of Hand Sanitizer Gel From Clove Flower Extract (Eugenia aromatica L.). International Journal of Science, Technology & Management, 3(2):484-491. https://doi.org/10.46729/ijstm.v3i2.472
Saheed, I. O., Da Oh, W., & Suah, F. B. M. (2021). Chitosan Modifications for Adsorption of Pollutants–a Review. Journal of Hazardous Materials, 408(15):1-61.
Saputra, D., Jamilah, J., & Lahay, N. (2023). Antibacterial Inhibition Test of Butterfly Pea (Clitoria ternatea L.) as Alternative Feed Additive for Broiler. AIP Conference Proceedings.
Somwanshi, S. B., Kumar, S., & Rathore, G. S. (2023). Cosmetic Science. (1st ed) AG Publishing House (AGPH Books). 241 p.
Sudianto., Suseno, S. H., & Suptijah, P. (2020). Optimization of Water-Soluble Chitosan Production Using the Pressure Hydrolysis Method. Jurnal Pengolahan Hasil Perikanan Indonesia, 23(3):441-446.
Wang, W., Xue, C., & Mao, X. (2020). Chitosan: Structural Modification, Biological Activity and Application. International Journal of Biological Macromolecules, 164(2020):4532-4546.
Wang, J., & Zhuang, S. (2022). Chitosan-based Materials: Preparation, Modification and Application. Journal of Cleaner Production, 355(2022):1- 25.
Wulandari, I. O., Pebriatin, B. E., Valiana, V., Hadisaputra, S., Ananto, A. D., & Sabarudin, A. (2022). Green Synthesis of Silver Nanoparticles Coated by Water Soluble Chitosan and its Potency as Non-alcoholic Hand Sanitizer Formulation. Materials, 15(13):1-20.
Zhou, J., Cai, Y., Liu, Y., An, H., Deng, K., Ashraf, M. A., Zou, L., & Wang, J. (2022). Breaking Down the Cell Wall: Still an Attractive Antibacterial Strategy. Frontiers in Microbiology, 13(2022):1-21.