Organic Acid-Based Natural Deep Eutectic Solvents: A Comparative Study for Chitin Extraction from Crab Shell By-Products
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Abstract
Conventional chitin extraction using acid/alkali method is effective but involves harsh chemicals. Biological methods offer improved consistency but often result in limited chitin solubility. This study evaluated the efficiency of lactic acid-based and succinic acid-based natural deep eutectic solvents (NADES) for chitin extraction from crab shells. Choline chloride was mixed with either lactic acid (lactic-NADES) or succinic acid (succinic-NADES) and combined with crab shell powder (CS) in a 10:1 ratio. The mixture was then heated using microwaves for varying durations. The results showed that lactic-NADES achieved higher demineralization and deproteinization efficiencies than succinic-NADES. Optimal extraction was achieved with 3 M lactic acid at a CS:lactic-NADES ratio of 1:20, with a microwave irradiation time of 5 min, yielding demineralization and deproteinization efficiencies of 97.78±0.73% and 81.33±0.91%, respectively. The microstructure, crystallographic organization, functional groups, and thermal stability of the optimal extracted chitin were then compared to those of conventionally extracted chitin. Analysis using SEM, FT-IR, XRD, and TGA showed that chitin obtained via the lactic-NADES method had characteristics comparable to those extracted using traditional acid/alkali methods. Thus, the lactic-NADES method presents a promising alternative for chitin extraction, offering similar quality with potentially fewer environmental impacts.
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References
Aranaz, I., M. Mengibar, R. Harris, I. Panos, B. Miralles, N. Acosta, G. Galed and A. Heras. 2009. Functional characterization of chitin and chitosan. Current Chemical Biology 3(2): 203–230. DOI: 10.2174/187231309788166415.
Dai, Y., J. van Spronsen, G.J. Witkamp, R. Verpoorte and Y.H. Choi. 2013a. Ionic liquids and deep eutectic solvents in natural products research: mixtures of solids as extraction solvents. Journal of Natural Products 76(11): 2162–2173. DOI: 10.1021/np400051w.
Dai, Y., J. van Spronsen, G.J. Witkamp, R. Verpoorte and Y.H. Choi. 2013b. Natural deep eutectic solvents as new potential media for green technology. Analytica Chimica Acta 766: 61–68. DOI: 10.1016/j.aca.2012.12.019.
de Queiroz Antonino, R., B.R.P. Lia Fook, V.A. de Oliveira Lima, R.I. de Farias Rached, E.P.N. Lima, R.J. da Silva Lima, C.O. Peniche Covas and M.V. Lia Fook. 2017. Preparation and characterization of chitosan obtained from shells of shrimp (Litopenaeus vannamei Boone). Marine Drugs 15(5): DOI: 10.3390/md15050141.
El Knidri, H., R. Belaabed, A. Addaou, A. Laajeb and A. Lahsini. 2018. Extraction, chemical modification and characterization of chitin and chitosan. International Journal of Biological Macromolecules 120: 1181–1189. DOI: 10.1016/j.ijbiomac.2018.08.139.
Gómez, A., A.Biswas, C. Tadini, R. Furtado, C. Alves and H. Cheng. 2019. Use of natural deep eutectic solvents for polymerization and polymer reactions. Journal of the Brazilian Chemical Society 30: 717–726. DOI: 10.21577/0103-5053.20190001.
Gomez, F.J.V., M. Espino, M.A. Fernández and M.F. Silva. 2018. A greener approach to prepare natural deep eutectic solvents. ChemistrySelect 3: 6122–6125. DOI: 10.1002/slct.201800713.
Gontrani, L., N.V. Plechkova and M. Bonomo. 2019. In-depth physico-chemical and structural investigation of a dicarboxylic acid/choline chloride natural deep eutectic solvent (NADES): A spotlight on the importance of a rigorous preparation procedure. ACS Sustainable Chemistry and Engineering 7(14): 12536–12543. DOI: 10.1021/acssuschemeng.9b02402.
Hamed, I., F. Özogul and J.M. Regenstein. 2016. Industrial applications of crustacean by-products (chitin, chitosan, and chitooligosaccharides): A review. Trends in Food Science and Technology 48: 40–50. DOI: 10.1016/j.tifs.2015.11.007.
Joseph, S.M., S. Krishnamoorthy, R. Paranthaman, J.A. Moses and C. Anandharamakrishnan. 2021. A review on source-specific chemistry, functionality, and applications of chitin and chitosan. Carbohydrate Polymer Technologies and Applications 2: 100036 DOI: 10.1016/j.carpta.2021.100036.
Kumari, S. and R. Kishor. 2020. Chitin and chitosan. In: Chitin and Chitosan: Origin, Properties, and Applications (eds. S. Gopi, S. Thomas and A. Pius), pp. 1–33. Elsevier, Bhopal, India.
Li, Z., M.C. Li, C. Liu, X. Liu, Y. Lu, G. Zhou, C. Liu and C. Mei. 2023. Microwave-assisted deep eutectic solvent extraction of chitin from crayfish shell wastes for 3D printable inks. Industrial Crops and Products 194: 116325. DOI: 10.1016/j.indcrop.2023.116325.
Liu, Y., J.B. Friesen, J.B. McAlpine, D.C. Lankin, S.N. Chen and G.F. Pauli. 2018. Natural deep eutectic solvents: Properties, applications, and perspectives. Journal of Natural Products 81(3): 679–690. DOI: 10.1021/acs.jnatprod.7b00945.
Margariti, C. 2019. The application of FTIR microspectroscopy in a non-invasive and non-destructive way to the study and conservation of mineralised excavated textiles. Heritage Science 7: 63. DOI: 10.1186/s40494-019-0304-8.
Ozel, N. and M. Elibol. 2021. A review on the potential uses of deep eutectic solvents in chitin and chitosan related processes. Carbohydrate Polymers 262: 117942. DOI: 10.1016/j.carbpol.2021.117942.
Pachapur, V.L., K. Guemiza, T. Rouissi, S.J. Sarma and S.K. Brar. 2016. Novel biological and chemical methods of chitin extraction from crustacean waste using saline water. Journal of Chemical Technology and Biotechnology 91: 2331–2339. DOI: 10.1002/jctb.4821.
Paiva, A., R. Craveiro, I. Aroso, M. Martins, R.L. Reis and A.R.C. Duarte. 2014. Natural deep eutectic solvents–Solvents for the 21st century. ACS Sustainable Chemistry and Engineering 2(5): 1063–1071. DOI: 10.1021/sc500096j.
Pedrol, N. and P. Tamayo. 2001. Plant ecophysiology techniques. In: Protein Content Quantification by Bradford Method (ed. M.J. Reigosa Roger), pp. 283–295. Kluwer Academic Publishers, Dordrecht, Netherlands.
Pemberton, A.T., D.B. Magers and D.A. King. 2018. Integrated TGA, FTIR, and computational laboratory experiment. Journal of Chemical Education 96(1): 132–136. DOI: 10.1021/acs.jchemed.8b00607.
Rahayu, A.P., A.F. Islami, E. Saputra, L. Sulmartiwi, A.U. Rahmah and K.A. Kurnia. 2022. The impact of the different types of acid solution on the extraction and adsorption performance of chitin from shrimp shell waste. International Journal of Biological Macromolecules 194: 843–850. DOI: 10.1016/j.ijbiomac.2021.11.137.
Santana, A.P.R., J.A. Mora-Vargas, T.G.S. Guimarães, C.D.B. Amaral, A. Oliveira and M.H. Gonzalez. 2019. Sustainable synthesis of natural deep eutectic solvents (NADES) by different methods. Journal of Molecular Liquids 293: 111452 DOI: 10.1016/j.molliq.2019.111452.
Saravana, P.S., T.C. Ho, S.J. Chae, Y.J. Cho, J.S. Park, H.J. Lee and B.S. Chun. 2018. Deep eutectic solvent-based extraction and fabrication of chitin films from crustacean waste. Carbohydrate Polymers 195: 622–630. DOI: 10.1016/j.carbpol.2018.05.018.
Saxena, R.K., S. Saran, J. Isar and R. Kaushik. 2017. Production and applications of succinic acid. In: Current Developments in Biotechnology and Bioengineering (eds. A. Pandey, S. Negi and C.R. Soccol), pp. 601–630. Elsevier, New delhi, India
Shamshina, J.L., P. Berton and R.D. Rogers. 2019. Advances in functional chitin materials: A review. ACS Sustainable Chemistry and Engineering 7(7): 6444–6457. DOI: 10.1021/acssuschemeng.8b06372.
Tolesa, L.D., B.S. Gupta and M.J. Lee. 2019. Chitin and chitosan production from shrimp shells using ammonium-based ionic liquids. International Journal of Biological Macromolecules 130: 818–826. DOI: 10.1016/j.ijbiomac.2019.03.018.
Vanda, H., Y. Dai, E.G. Wilson, R. Verpoorte and Y.H. Choi. 2018. Green solvents from ionic liquids and deep eutectic solvents to natural deep eutectic solvents. Comptes Rendus Chimie 21(6): 628–638. DOI: 10.1016/j.crci.2018.04.002.
Vicente, F.A., M. Hus, B. Likozar and U. Novak. 2021. Chitin deacetylation using deep eutectic solvents: Ab initio-supported process optimization. ACS Sustainable Chemistry and Engineering 9(10): 3874–3886. DOI: 10.1021/acssuschemeng.0c08976.
Yang, Z. 2019. Natural deep eutectic solvents and their applications in biotechnology. Advances in Biochemical Engineering/Biotechnology 168: 31–59. DOI: 10.1007/10_2018_67.
Zhu, P., Z. Gu, S. Hong and H. Lian. 2017. One-pot production of chitin with high purity from lobster shells using choline chloride-malonic acid deep eutectic solvent. Carbohydrate Polymers 177: 217–223. DOI: 10.1016/j.carbpol.2017.09.001.