Influence of CaCl2 Concentration on the physicochemical and structural properties of crude fucoidan from Sargassum sp.: Pengaruh konsentrasi CaCl2 terhadap karakteristik fisikokimia dan struktural fukoidan kasar Sargassum sp.

Ulfah  Amalia; Widan Adi  Darmawan; Siti Susanti; Ima  Wijayanti

doi:10.17844/8srd9c23

Authors

Ulfah Amalia Department of Fisheries Product Technology, Faculty of Fisheries and Marine Science, Diponegoro University https://orcid.org/0000-0002-5707-6079
Widan Adi Darmawan Department of Fisheries Product Technology, Faculty of Fisheries and Marine Science, Diponegoro University
Siti Susanti Food Technology, Department of Agriculture, Faculty of Animal and Agricultural Sciences, Diponegoro University https://orcid.org/0000-0001-6046-0381
Ima Wijayanti Department of Fisheries Product Technology, Faculty of Fisheries and Marine Science, Diponegoro University https://orcid.org/0000-0002-7082-1424

DOI:

https://doi.org/10.17844/8srd9c23

Keywords:

brown seaweed , color, infrared spectroscopy, sulfated polysaccharide, yield

Abstract

Fucoidan is a sulfated polysaccharide derived from brown seaweed. Fucoidan exhibits various biological activities and potential therapeutic applications. The extraction method and conditions determine the yield and characteristics of the fucoidan products. This study aimed to evaluate the physicochemical characteristics and structure of crude fucoidan extracted using various concentrations of CaCl₂. Crude fucoidan was extracted using CaCl₂ solutions with concentrations of 2, 3, and 4 M. Parameters analyzed included yield, moisture, ash, pH, color, and functional groups using Fourier Transform Infrared Spectroscopy (FT-IR). The results showed that extraction using 2 M CaCl₂ produced the highest yield (8%) and moisture (13.3%). Extraction using 4 M CaCl₂ showed the highest absorption intensity for sulfate and hydroxyl groups, as indicated by the FT-IR spectrum. The ash content did not differ significantly between the treatments and ranged from 0.50-0.57%. The pH of the extract was in the neutral range, namely 7.01-7.11. Color analysis showed that increasing the CaCl ₂ concentration produced a greenish-brown color. Extraction using 3 M CaCl₂ resulted in the highest brightness (L* = 25.31) and lowest redness (a* = −0.56). FT-IR analysis confirmed the presence of hydroxyl, carbonyl, sulfate ester, and glycosidic bonds as characteristic features of the fucoidan structure. This study concluded that the concentration of CaCl₂ affects the yield and structural characteristics of crude fucoidan. The selection of CaCl₂ concentration needs to be adjusted to the utilization objectives, both to obtain high yields and to improve the characteristics of sulfate groups that have the potential to support bioactivity.

References

Alboofetileh, M., Tabarsa, M., & Rezaei, M. (2018). Enzyme-assisted extraction of Nizamuddinia zanardinii for the recovery of sulfated polysaccharides with anticancer and immune-enhancing activities. Journal of Applied Phycology, 31(2), 1391–1402. https://doi.org/10.1007/s10811-018-1651-7

Ale, M. T., Mikkelsen, J. D., & Meyer, A. S. (2011). Important determinants for fucoidan bioactivity: A critical review of structure-function relations and extraction methods for fucose-containing sulfated polysaccharides from brown seaweeds. Marine Drugs, 9(10), 2106–2130 https://doi.org/10.3390/md9102106

Ale, M. T., Mikkelsen, J. D., & Meyer, A. S. (2012). Designed optimization of a single-step extraction of fucose-containing sulfated polysaccharides from Sargassum sp. Journal of Applied Phycology, 24(4), 715–723. https://doi.org/10.1007/s10811-011-9690-3

AOAC. (2005). Official methods of analysis of the association of official analytical chemists international. Maryland.

Brown, T. L., LeMay, H. E., Bursten, B. E., Murphy, C. J., & Woodward, P. (2018). Chemistry: The Central Science (14th ed.). Pearson Education.

Chadwick M, Carvalho LG, Vanegas C, Dimartino S. (2025). A comparative review of alternative fucoidan extraction techniques from seaweed. Marine Drugs, 23(1), 1-36. https://doi.org/10.3390/md23010027

Chang, R., & Goldsby, K. A. (2016). Chemistry (12th ed.). McGraw-Hill Education.

Choi, J. il, Gu Lee, S., Jong Han, S., Cho, M., & Cheon Lee, P. (2014). Effect of gamma irradiation on the structure of fucoidan. Radiation Physics and Chemistry, 100, 54–58. https://doi.org/10.1016/j.radphyschem.2014.03.018

Cugmas, B., & Štruc, E. (2020). Accuracy of an affordable smartphone-based teledermoscopy system for color measurements in canine skin. Sensors, 20(21), 1-12. https://doi.org/10.3390/s20216234

Dhahri, M. (2023). Cystoseira myrica: from beach-cast seaweed to fucoidan with antioxidant and anticoagulant capacity. Frontiers in Marine Science, 10, 1-11. https://doi.org/10.3389/fmars.2023.1327408

Dörschmann, P., Bittkau, K. S., Neupane, S., Roider, J., Alban, S., & Klettner, A. (2019). Effects of fucoidans from five different brown algae on oxidative stress and VEGF interference in ocular cells. Marine Drugs, 17(5), 1-19. https://doi.org/10.3390/md17050258

El-Sheekh, M., Alwaleed, E. A., Kassem, W. M. A., & Saber, H. (2024). Optimizing the fucoidan extraction using Box-Behnken Design and its potential bioactivity. International Journal of Biological Macromolecules, 277. https://doi.org/10.1016/j.ijbiomac.2024.134490

Fernando, S., I. P., Abeytunga, D. T. U., Samarakoon, K. W., Asanka Sanjeewa, K. K., Gunasekara, U. K. D. S. S., Jeon, Y.-J., Lee, W. W., Kim, H.-S., & Park, Y.-J. (2018). The potential of fucoidans from Chnoospora minima and Sargassum polycystum in cosmetics: antioxidant, anti-inflammatory, skin-whitening, and antiwrinkle activities. Journal of Applied Phycology, 30(6), 3223–3232. https://doi.org/10.1007/s10811-018-1415-4

Flórez-Fernández, N., González-Muñoz, M. J., López-García, M., Domínguez, H., & Vilariño, J. M. L. (2017). Ultrasound-assisted extraction of fucoidan from Sargassum muticum. Journal of Applied Phycology, 29(3), 1553–1561. https://doi.org/10.1007/s10811-016-1043-9

George, A., & Shrivastav, P. S. (2023). Fucoidan, a brown seaweed polysaccharide in nanodrug delivery. Drug Delivery and Translational Research, 13(10), 2427–2446. https://doi.org/10.1007/s13346-023-01329-4

Goñi, S. M., & Salvadori, V. O. (2016). Color measurement: comparison of colorimeter vs. computer vision system. Journal of Food Measurement and Characterization, 11(2), 538–547. https://doi.org/10.1007/s11694-016-9421-1

Hifney, A. F., Fawzy, M. A., Abdel-Gawad, K. M., & Gomaa, M. (2016). Industrial optimization of fucoidan extraction from Sargassum sp. and its potential antioxidant and emulsifying activities. Food Hydrocolloids, 54, 77–88. https://doi.org/10.1016/j.foodhyd.2015.09.022

Husni, A., Husnayain, N., Ayunani, F. Z., Izmi, N., Kartini, A., & Isnansetyo, A. (2022). Characteristics and antioxidant activity of fucoidan from Sargassum hystrix: effect of extraction method. International Journal of Food Science, 1-9 https://doi.org/10.1155/2022/3689724

Imbs, T. I., Skriptsova, A. V., & Zvyagintseva, T. N. (2014) Antioxidant activity of fucosecontaining sulfated polysaccharides obtained from Fucus evanescens by different extraction methods. Journal of Applied Phycology 27, 545-553. https://doi.org/10.1007/s10811-014-0293-7

Isnansetyo, A., Murwantoko, M., Kasanah, N., & Fikriyah, A. (2015). Non-specific immune potentiating activity of fucoidan from a tropical brown algae (Phaeophyceae), Sargassum cristaefolium in tilapia (Oreochromis niloticus). Aquaculture International, 24(2), 465–477. https://doi.org/10.1007/s10499-015-9938-z

Jeong, S., Lee, S., Lee, G., Hyun, J., & Ryu, B. (2024). Systematic characteristics of fucoidan: intriguing features for new pharmacological interventions. International Journal of Molecular Sciences, 25(21), 1-21. https://doi.org/10.3390/ijms252111771

Joung, E. J., Choi, J., Jung, B. M., Kim, H. R., Gwon, W. G., & Shin, T. (2016). Anti-inflammatory action of the ethanolic extract from Sargassum serratifolium on lipopolysaccharide-stimulated mouse peritoneal macrophages and identification of active components. Journal of Applied Phycology, 29(1), 563–573. https://doi.org/10.1007/s10811-016-0954-9

Kang, J. W., Lee, I. C., Bae, J. S., Hyun, S. H., Kim, H. M., Lee, J. A., & Park, S. Y. (2023). The effects of fucoidan-rich polysaccharides extracted from Sargassum horneri on enhancing collagen-related skin barrier function as a potential cosmetic product. Journal of Cosmetic Dermatology, 23(4), 1365–1373. https://doi.org/10.1111/jocd.16108

Lee, M. K., Kim, M. J., Lee, B., Baek, J., Lee, J. Y., Ryu, H., Van, J. Y., Jung, W. K., & Jeong, H. H. (2022). Potential beneficial effects of Sargassum spp. in skin aging. Marine Drugs, 20(8), 1-17. https://doi.org/10.3390/md20080540

Li, B., Lu, F., Wei, X., & Zhao, R. (2008). Fucoidan: structure and bioactivity. Molecules, 13(8), 1671–1695. https://doi.org/10.3390/molecules13081671

Lin, Y., Xu, S., Tian, Z., Liu, H., Xue, K., & Qi, X. (2020). The anti-cancer effects of fucoidan: a review of both in vivo and in vitro investigations. Cancer Cell International, 20(1), 1-14. https://doi.org/10.1186/s12935-020-01233-8

Liu, J., Tong, H., Choi, J. I., Luthuli, S., Wu, M., & Wu, Q. (2020). Pharmaceutical and nutraceutical potential applications of Sargassum fulvellum. BioMed Research International, 2020(6), 1–12. https://doi.org/10.1155/2020/2417410

Lorbeer, A. J., Zhang, W., Fincher, G. B., Lahnstein, J., & Su, P. (2014). Kinetics of conventional and microwave-assisted fucoidan extractions from the brown alga, Ecklonia radiata. Journal of Applied Phycology, 27(5), 2079–2087. https://doi.org/10.1007/s10811-014-0446-8

Mabate, B., Daub, C. D., Malgas, S., Edkins, A. L., & Pletschke, B. I. (2021). Fucoidan structure and its impact on glucose metabolism: Implications for diabetes and cancer therapy. Marine Drugs, 19(1), 1-20. https://doi.org/10.3390/md19010030

Manteu, S. H., Nurjanah, Abdullah, A., Nurhayati, T., & Seulalae, A. V. (2021). Efektivitas karbon aktif dalam pembuatan garam rumput laut cokelat (Sargassum polycystum dan Padina minor). Jurnal Pengolahan Hasil Perikanan Indonesia, 24(3), 407-416. https://doi.org/10.17844/jphpi.v24i3.26692

Nguyen, T. T., Mikkelsen, M. D., Tran, V.H.N., Trang, V.T.D., Rhein-Knudsen, N., Holck, J., Rasin, A. B., Cao, H. T. T., Van, T.T.T., & Meyer, A. S. (2020). Enzyme-Assisted fucoidan extraction from brown macroalgae Fucus distichus subsp. evanescent and Saccharina latissimia. Marine Drugs, 18(6), 1-18. https://doi.org/10.3390/md18060296

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), 1-10. https://doi.org/10.3390/jmse8060440

Nurhidayati, L., Fitriani, Y., Abdillah, S., Mumpuni, E., & Rafi, M. (2020). Sifat fisikokimia dan aktivitas antioksidan crude fukoidan hasil ekstraksi dari Sargassum cinereum. Jurnal Ilmu Kefarmasian Indonesia, 18(1), 68–74. https://doi.org/ 10.35814/jifi.v18i1.823

Oh, J. Y., Kim, E. A., Kang, S. I., Yang, H. W., Ryu, B., Wang, L., Lee, J. S., & Jeon, Y. J. (2020). Protective effects of fucoidan isolated from celluclast-assisted extract of Undaria pinnatifida sporophylls against aaph-induced oxidative stress in vitro and in vivo zebrafish model. Molecules (Basel, Switzerland), 25(10), 1-11. https://doi.org/10.3390/molecules25102361

Oliyaei, N., & Moosavi‐Nasab, M. (2021). Ultrasound‐assisted extraction of fucoxanthin from Sargassum angustifolium and Cystoseira indica brown algae. Journal of Food Processing and Preservation, 45(11), 1-10. https://doi.org/10.1111/jfpp.15929

Puspantari, W., Kusnandar, F., Lioe, N. H., & Laily, N. (2020). Penghambatan fraksi fukoidan rumput laut cokelat (Sargassum polycystum dan Turbinaria conoides) terhadap α-Amilase dan α-Glukosidase. Jurnal Pengolahan Hasil Perikanan Indonesia, 23(1), 122–136. https://doi.org/10.17844/jphpi.v23i1.30925

Rajauria, G., Ravindran, R., Garcia-Vaquero, M., Rai, D. K., Sweeney, T., & O’Doherty, J. (2023). Purification and molecular characterization of fucoidan isolated from Ascophyllum nodosum brown seaweed grown in Ireland. Marine Drugs, 21(5), 1-15. https://doi.org/10.3390/md21050315

Ramu, S., Murali, A., Narasimhaiah, G., & Jayaraman, A. (2020). Toxicological evaluation of Sargassum Wightii greville derived fucoidan in wistar rats: Haematological, biochemical and histopathological evidences. Toxicology Reports, 7, 874–882. https://doi.org/10.1016/j.toxrep.2020.07.009

Rodrigues, D., Rocha-Santos, T. A. P., Silva, A., Freitas, A. C., Duarte, A. C., Sousa, S., Pereira, L., Gomes, A. M. P., & Amorim, M. (2015). Impact of enzyme- and ultrasound-assisted extraction methods on biological properties of red, brown, and green seaweeds from the central west coast of Portugal. Journal of Agricultural and Food Chemistry, 63(12), 3177–3188. https://doi.org/10.1021/jf504220e

Rushdi, M. I., Saber, H., Hassan, H. M., Abdelmohsen, U. R., Elmaidomy, A. H., Abdelraheem, W. M., Madkour, H. A., Attia, E. Z., & Abdel-Rahman, I. A. M. (2020). Pharmacological and natural products diversity of the brown algae genus Sargassum. RSC Advances, 10(42), 24951–24972. https://doi.org/10.1039/d0ra03576a

Saeed, M., Noreldin, A. E., Elnesr, S. S., Dhama, K., Umar, M., Marghazani, I. B., Bhutto, Z. A., Soomro, F., Arain, M. A., Chao, S., Abd El‐Hack, M. E., Alagawany, M., Farag, M. R., Ali Fazlani, S., & Soomro, J. (2021). A comprehensive review on the health benefits and nutritional significance of fucoidan polysaccharide derived from brown seaweeds in human, animals and aquatic organisms. Aquaculture Nutrition, 27(3), 633–654. https://doi.org/10.1111/anu.13233

Salehi, B., Sharifi-Rad, J., Seca, A. M. L., Pinto, D. C. G. A., Michalak, I., Trincone, A., Mishra, A. P., Nigam, M., Zam, W., & Martins, N. (2019). Current trends on seaweeds: Looking at chemical composition, phytopharmacology, and cosmetic applications. IMolecules 24(22), 1-50. https://doi.org/10.3390/molecules24224182

Sanniyasi, E., Venkatasubramanian, G., Anbalagan, M. M., Raj, P. P., & Gopal, R. K. (2019). In vitro anti-HIV-1 activity of the bioactive compound extracted and purified from two different marine macroalgae (seaweeds) (Dictyota bartayesiana J.V.Lamourouxxa0and Turbinaria decurrensxa0Bory). Scientific Reports, 9(1), 1-12. https://doi.org/10.1038/s41598-019-47917-8

Ummat, V., Sivagnanam, S. P., Rai, D. K., O’Donnell, C., Conway, G. E., Heffernan, S. M., Fitzpatrick, S., Lyons, H., Curtin, J., & Tiwari, B. K. (2024). Conventional extraction of fucoidan from Irish brown seaweed Fucus vesiculosus followed by ultrasound-assisted depolymerization. Scientific Reports, 14(1), 1-13. https://doi.org/10.1038/s41598-024-55225-z

Wang, J., Zhang, Q., Zhang, Z., Song, H., & Li, P. (2010). Potential antioxidant and anticoagulant capacity of low molecular weight fucoidan fractions extracted from Laminaria japonica. International Journal of Biological Macromolecules, 46(1), 6–12. https://doi.org/10.1016/j.ijbiomac.2009.10.015

Xing, R., Liu, S., Yu, H., Chen, X., Qin, Y., Li, K., & Li, P. (2013). Extraction and separation of fucoidan from Laminaria japonica with chitosan as extractant. BioMed Research International, 1-4. https://doi.org/10.1155/2013/193689

Yende, S., Harle, U., & Chaugule, B. (2014). Therapeutic potential and health benefits of Sargassum species. Pharmacognosy Reviews, 8(15), 1-7. https://doi.org/10.4103/0973-7847.125514

Yu, J., White, W. L., Zhu, W., Liu, Y., Zhao, Y., Tang, W., Nie, S., Wu, J., Li, Q., Lu, J., Yang, X., Yang, S., & Hassouna, A. (2021). Fucoidan extracted from Sporophyll of Undaria pinnatifida Grown in Weihai, China - chemical composition and comparison of antioxidant activity of different molecular weight fractions. Frontiers in Nutrition, 8, 1-11. https://doi.org/10.3389/fnut.2021.636930

Zayed, A., Rupp, S., Burger-Kentischer, A., Ulber, R., Muffler, K., Hahn, T., & Finkelmeier, D. (2016). Physicochemical and biological characterization of fucoidan from Fucus vesiculosus purified by dye affinity chromatography. Marine Drugs, 14(4), 1-15. https://doi.org/10.3390/md14040079