ICOPMAP SPECIAL EDITION Optimisation of sulfate polysaccharide extraction from Ulva lactuca using natural deep eutectic solvent with ultrasound assistance
Heru Mukti1 , Abdul Mun’im2 , Fadlina Chany Saputri3 , Masteria Yunovilsa Putra4
DOI:
https://doi.org/10.46542/pe.2025.252.6876Abstract
Background: Ulva lactuca, or sea lettuce, is a green macroalgae found in the Indonesian Sea, containing the active component Ulvan sulfated polysaccharide.
Objective: This study aims to investigate the impact of extraction-free variables on ulvan yield and its corresponding metabolomic profile.
Methods: Ulvan was extracted using three choline chloride-based NADES (KKG, KKL, KKT) with ultrasonic waves, optimising variables such as temperature, solution-to-solid ratio, and power. Total polysaccharides were measured using the phenol-sulfuric acid method, while total sulfates were measured by BaCl2 absorption turbidimetry at 490 nm and 420 nm. Ulvan metabolomic profiles were assessed via FTIR, XRD, and LC-HRMS.
Results: The independent variables consisted of 3 factors and 3 levels, resulting in 51 test points with optimum parameters: temperature at 78.27°C, L/P ratio at 39.62, and ultrasonic power at 139.23 watts. KKG achieved the highest yield at 11.7% ± 0.706, with sulphate levels ranging from 6.37% to 10.4% across all NADES. Extraction results were influenced by both single and combined independent variables. FTIR metabolomics revealed -CH, -S=O, -C=O, and -SO₄ groups, while LC-HRMS identified Rhamnosa 3-sulphate and DI-Xylosa compounds. UHD contains the Xylose-Rhamnosa 3-sulphate chain, part of Ulvanobiosis U3s.
Conclusion: Choline chloride-based NADES, with ultrasound assistance, shows potential as a method for isolating active compounds from marine biota.
References
Abdallah, M. M., Cardeira, M., Matias, A. A., Bronze, M. R., & Fernández, N. (2022). Lactic acid-based natural deep eutectic solvents to extract bioactives from marine by-products. Molecules, 27(14), 1–14. https://doi.org/10.3390/molecules27144356
Ahmad, A., Rehman, M. U., Wali, A. F., El-Serehy, H. A., Al-Misned, F. A., Maodaa, S. N., Aljawdah, H. M., Mir, T. M., & Ahmad, P. (2020). Box–Behnken response surface design of polysaccharide extraction from Rhododendron arboreum and the evaluation of its antioxidant potential. Molecules, 25(17), 3835. https://doi.org/10.3390/molecules25173835
Andryukov, B. G., Besednova, N. N., Kuznetsova, T. A., Zaporozhets, T. S., Ermakova, S. P., Zvyagintseva, T. N., Chingizova, E. A., Gazha, A. K., & Smolina, T. P. (2020). Sulfated polysaccharides from marine algae as a basis of modern biotechnologies for creating wound dressings: Current achievements and future prospects. Biomedicines, 8(9). https://doi.org/10.3390/biomedicines8090301
Barakat, K. M., Ismail, M. M., Abou El Hassayeb, H. E., El Sersy, N. A., & Elshobary, M. E. (2022). Chemical characterization and biological activities of ulvan extracted from Ulva fasciata (Chlorophyta). Rendiconti Lincei, 33(4), 829–841. https://doi.org/10.1007/s12210-022-01103-7
Carvalho, D. N., Inácio, A. R., Sousa, R. O., Reis, R. L., & Silva, T. H. (2020). Seaweed polysaccharides as sustainable building blocks for biomaterials in tissue engineering. In Sustainable seaweed technologies: Cultivation, biorefinery, and applications. Elsevier Inc. https://doi.org/10.1016/B978-0-12-817943-7.00019-6
Chakraborty, K., Joseph, D., & Praveen, N. K. (2015). Antioxidant activities and phenolic contents of three red seaweeds (Division: Rhodophyta) harvested from the Gulf of Mannar of Peninsular India. Journal of Food Science and Technology, 52(4), 1924–1935. https://doi.org/10.1007/s13197-013-1189-2
Chi, Y., Li, H., Wang, P., Du, C., Ye, H., Zuo, S., Guan, H., & Wang, P. (2020). Structural characterization of ulvan extracted from Ulva clathrata assisted by an ulvan lyase. Carbohydrate Polymers, 229, 115497. https://doi.org/10.1016/j.carbpol.2019.115497
Dhahri, M., Sioud, S., Dridi, R., Hassine, M., Boughattas, N. A., Almulhim, F., Al Talla, Z., Jaremko, M., & Emwas, A. H. M. (2020). Extraction, characterization, and anticoagulant activity of a sulfated polysaccharide from Bursatella leachii Viscera. ACS Omega, 5(24), 14786–14795. https://doi.org/10.1021/acsomega.0c01724
Dubois, M., Gilles, K., Hamilton, J. K., Rebers, P. A., & Smith, F. (1951). A colorimetric method for the determination of sugars. Nature, 168(4265), 167. https://doi.org/10.1038/168167a0
Fanali, C., Gallo, V., Posta, S. Della, Dugo, L., Mazzeo, L., Cocchi, M., Piemonte, V., & De Gara, L. (2021). Choline chloride–lactic acid-based NADES as an extraction medium in a response surface methodology-optimized method for the extraction of phenolic compounds from hazelnut skin. Molecules, 26(9), 2652. https://doi.org/10.3390/molecules26092652
Figueroa, F. A., Abdala-d, R. T., Claudia, P., Casas-arrojo, V., Nesic, A., Tapia, C., Dur, C., Valdes, O., Parra, C., Bravo-arrepol, G., Soto, L., Becerra, J., & Cabrera-barjas, G. (2022). Sulfated polysaccharide extracted from the green algae Codium bernabei: physicochemical characterization and antioxidant, anticoagulant and antitumor activity. Marine Drugs, 20(7), 458. https://doi.org/10.3390/md20070458
Georgiopoulou, I., Louli, V., & Magoulas, K. (2023). Comparative study of conventional, microwave-assisted and supercritical fluid extraction of bioactive compounds from microalgae: the case of Scenedesmus obliquus. Separations, 10(5). https://doi.org/10.3390/separations10050290
Hung, Y. H. R., Chen, G. W., Pan, C. L., & Lin, H. T. V. (2021). Production of ulvan oligosaccharides with antioxidant and angiotensin-converting enzyme-inhibitory activities by microbial enzymatic hydrolysis. Fermentation, 7(3), 160. https://doi.org/10.3390/fermentation7030160
Jeliński, T., Przybyłek, M., & Cysewski, P. (2019). Natural deep eutectic solvents as agents for improving solubility, stability and delivery of curcumin. Pharmaceutical Research, 36(8), 116. https://doi.org/10.1007/s11095-019-2643-2
Karaosmanoglu, S., Zhou, M., Shi, B., Zhang, X., Williams, G. R., & Chen, X. (2021). Carrier-free nanodrugs for safe and effective cancer treatment. Journal of Controlled Release, 329, 805–832. https://doi.org/10.1016/J.JCONREL.2020.10.014
Lahaye, M., & Robic, A. (2007). Structure and function properties of Ulvan, a polysaccharide from green seaweeds. Biomacromolecules, 8(6), 1765–1774. https://doi.org/10.1021/bm061185q
Ling, J. K. U., & Hadinoto, K. (2022). Deep eutectic solvent as green solvent in extraction of biological macromolecules: A Review. International Journal of Molecular Sciences, 23(6), 3381. https://doi.org/10.3390/ijms23063381
Liu, J.-Z., Lu, X.-X., Yang, X.-T., Jiang, L.-J., & Cui, Q. (2024). An efficient approach for the extraction of polyphenols from pomegranate peel using the green solvent and profiling by UPLC-Q-TOF-MS/MS analysis. Microchemical Journal, 205, 111421. https://doi.org/https://doi.org/10.1016/j.microc.2024.111421
Liu, J., Song, J., Gao, F., Chen, W., Zong, Y., Li, J., He, Z., & Du, R. (2023). Extraction, purification, and structural characterization of polysaccharides from Sanghuangporus vaninii with anti-inflammatory activity. Molecules, 28(16), 6081. https://doi.org/10.3390/molecules28166081
Lu, W., & Liu, S. (2022). Choline chloride–based deep eutectic solvents (Ch-DESs) as promising green solvents for phenolic compounds extraction from bioresources: State-of-the-art, prospects, and challenges. Biomass Conversion and Biorefinery, 12(7), 2949–2962. https://doi.org/10.1007/s13399-020-00753-7
Manikandan, N. A., & Lens, P. N. L. (2022). Green extraction and esterification of marine polysaccharide (ulvan) from green macroalgae Ulva sp. using citric acid for hydrogel preparation. Journal of Cleaner Production, 366, 132952. https://doi.org/10.1016/j.jclepro.2022.132952
Manukovskaya, M. V, Shchetilina, I. P., Pisklyukova, Y. N., Klimova, E. A., & Korystin, M. I. (2021). The use of ultrasonic extraction in the technology of functional drinks based on plant raw materials. IOP Conference Series: Earth and Environmental Science, 845(1), 12114. https://doi.org/10.1088/1755-1315/845/1/012114
Menezes Maciel Bindes, M., Hespanhol Miranda Reis, M., Luiz Cardoso, V., & Boffito, D. C. (2019). Ultrasound-assisted extraction of bioactive compounds from green tea leaves and clarification with natural coagulants (chitosan and Moringa oleífera seeds). Ultrasonics Sonochemistry, 51, 111–119. https://doi.org/https://doi.org/10.1016/j.ultsonch.2018.10.014
Nie, J., Chen, D., & Lu, Y. (2020). Deep eutectic solvents based ultrasonic extraction of polysaccharides from edible brown Seaweed Sargassum horneri. Journal of Marine Science and Engineering, 8(6), 440. https://doi.org/10.3390/JMSE8060440
Nipornram, S., Tochampa, W., Rattanatraiwong, P., & Singanusong, R. (2017). Optimization of low power ultrasound-assisted extraction of phenolic compounds from mandarin (Citrus Reticulata Blanco cv. Sainampueng) peel. Food Chemistry, 241, 338–345. https://doi.org/10.1016/j.foodchem.2017.08.114
Olsson, J., Toth, G. B., Oerbekke, A., Cvijetinovic, S., Wahlström, N., Harrysson, H., Steinhagen, S., Kinnby, A., White, J., Edlund, U., Undeland, I., Pavia, H., & Albers, E. (2020). Cultivation conditions affect the monosaccharide composition in Ulva fenestrata. Journal of Applied Phycology, 32(5), 3255–3263. https://doi.org/10.1007/s10811-020-02138-9
Qi, X.-L., Peng, X., Huang, Y.-Y., Li, L., Wei, Z.-F., Zu, Y.-G., & Fu, Y.-J. (2015). Green and efficient extraction of bioactive flavonoids from Equisetum palustre L. by deep eutectic solvents-based negative pressure cavitation method combined with macroporous resin enrichment. Industrial Crops and Products, 70, 142–148. https://doi.org/https://doi.org/10.1016/j.indcrop.2015.03.026
Robic, A., Gaillard, C., Sassi, J. F., Leral, Y., & Lahaye, M. (2009). Ultrastructure of ulvan: A polysaccharide from green seaweeds. Biopolymers, 91(8), 652–664. https://doi.org/10.1002/bip.21195
Şahin, S., Aybastıer, Ö., & Işık, E. (2013). Optimisation of ultrasonic-assisted extraction of antioxidant compounds from Artemisia absinthium using response surface methodology. Food Chemistry, 141(2), 1361–1368. https://doi.org/https://doi.org/10.1016/j.foodchem.2013.04.003
Sari-Chmayssem, N., Taha, S., Mawlawi, H., Guégan, J.-P., Jeftić, J., & Benvegnu, T. (2019). Extracted ulvans from green algae Ulva linza of Lebanese origin and amphiphilic derivatives: evaluation of their physico-chemical and rheological properties. Journal of Applied Phycology, 31. https://doi.org/10.1007/s10811-018-1668-y
Sun, Y., Zhang, M., & Fang, Z. (2020). Efficient physical extraction of active constituents from edible fungi and their potential bioactivities: A review. Trends in Food Science & Technology, 105, 468–482. https://doi.org/10.1016/J.TIFS.2019.02.026
Thanh, T. T. T., Quach, T. M. T., Nguyen, T. N., Vu Luong, D., Bui, M. L., & Tran, T. T. Van. (2016). Structure and cytotoxic activity of ulvan extracted from green seaweed Ulva lactuca. International Journal of Biological Macromolecules, 93, 695–702. https://doi.org/10.1016/j.ijbiomac.2016.09.040
Wang, M., Wang, J., Zhang, Y., Xia, Q., Bi, W., Yang, X., & Chen, D. D. Y. (2016). Fast environment-friendly ball mill-assisted deep eutectic solvent-based extraction of natural products. Journal of Chromatography A, 1443, 262–266. https://doi.org/https://doi.org/10.1016/j.chroma.2016.03.061