PENGARUH WAKTU DAN PELARUT EDTA (Ethylenediaminetetraacetic Acid) PADA EKSTRAKSI FUKOIDAN DARI RUMPUT LAUT COKELAT Sargassum binderi Sonder

Rumput laut cokelat memiliki beragam polisakarida yang terkandung di dalamnya. Salah satu polisakarida dari rumput laut cokelat yang diketahui memiliki bioaktivitas yang tinggi adalah fukoidan. Banyak penelitian telah menunjukkan bahwa bioaktivitas dari fukoidan antara lain mampu menghambat pertumbuhan sel kanker, anti koagulan, anti virial, dan sebagai immunostimulan. Mengisolasi fukoidan dari rumput laut cokelat telah banyak dilakukan dengan beragam cara. Penelitian ini bertujuan untuk mengetahui bagaimana pengaruh waktu dan konsentrasi pelarut EDTA (ethylenediaminetetraacetic acid) pada proses ekstraksi dan karakteristik fukoidan dari rumput laut cokelat jenis Sargassum binderi Sonder. Konsentrasi pelarut yang digunakan pada penelitian ini sebesar 0,15% ; 0,3%: 0,5%; 1%; dan 1,3% serta waktu ekstraksi selama 1 jam, 2 jam, 3 jam, 4 jam, dan 5 jam. Perbandingan rumput laut dan air adalah 1:30 dengan suhu ekstraksi dilakukan pada 70˚C. Dalam penelitian ini didapatkan yield crude fucoidan terbesar pada waktu ekstraksi 4 jam dan konsentrasi pelarut 1,3% yaitu sebesar 3,67%. Hasil analisis menunjukkan kandungan total gula dari ekstrak fukoidan dengan yield terbesar didapatkan sebesar 36,48% dan kandungan sulfat sebesar 17,16%.

Crude Fucoidan Activity Extracted from Sargassum sp. as Mycotoxin (T-2) Binder

Fucoidan is a sulfated polysaccharide which is mainly found in brown seaweed like Sargassum sp. In this study, fucoidan was extracted with two distinct methods, one was using 0.1 N HCl (Fucoidan A) and the other 0.1 N H3PO4 (Fucoidan B). The yield of HCl extraction was respectively 7.50% and H3PO4 extraction 7.02%. Characterization of crude fucoidan was carried out through fourier-transform infrared spectroscopy (FTIR), followed by total carbohydrate and total sulfate level measurement. Crude fucoidan was then used to determine its binding activity against Trichothecenes T-2 toxin. Quantitative analysis of 50 ug/ml T-2 toxin binding capacity shows an efficiency of 59.52% at pH 3.0 and 58.37% at pH 6.8 for crude fucoidan A. As for crude fucoidan B, efficiency of T-2 toxin binding capacity has value of 58.12% at pH 3.0 and 57.33% at pH 6.8. Meanwhile, Commercial crude fucoidan extract from Fucus vesiculosus sp. was used as control for T-2 toxin binding capacity analysis with efficiency of 57.31% at pH 3.0 and 56.64% at pH 6.8. Therefore, fucoidan from Sargassum sp. can be utilized better as mycotoxin especially T-2 toxin binder than from Fucus vesiculosus sp. through efficiency result of fucoidan binding capacity.

Fucoidan Inhibition of Osteosarcoma Cells is Species and Molecular Weight Dependent

Fucoidan is a brown algae-derived polysaccharide having several biomedical applications. This study simultaneously compares the anti-cancer activities of crude fucoidans from Fucus vesiculosus and Sargassum filipendula, and effects of low (LMW, 10–50 kDa), medium (MMW, 50–100 kDa) and high (HMW, >100 kDa) molecular weight fractions of S. filipendula fucoidan against osteosarcoma cells. Glucose, fucose and acid levels were lower and sulphation was higher in F. vesiculosus crude fucoidan compared to S. filipendula crude fucoidan. MMW had the highest levels of sugars, acids and sulphation among molecular weight fractions. There was a dose-dependent drop in focal adhesion formation and proliferation of cells for all fucoidan-types, but F. vesiculosus fucoidan and HMW had the strongest effects. G1-phase arrest was induced by F. vesiculosus fucoidan, MMW and HMW, however F. vesiculosus fucoidan treatment also caused accumulation in the sub-G1-phase. Mitochondrial damage occurred for all fucoidan-types, however F. vesiculosus fucoidan led to mitochondrial fragmentation. Annexin V/PI, TUNEL and cytochrome c staining confirmed stress-induced apoptosis-like cell death for F. vesiculosus fucoidan and features of stress-induced necrosis-like cell death for S. filipendula fucoidans. There was also variation in penetrability of different fucoidans inside the cell. These differences in anti-cancer activity of fucoidans are applicable for osteosarcoma treatment.

Fucoidan Inhibition of Osteosarcoma Cells is Species and Molecular Weight Dependent

Fucoidan is a brown algae-derived polysaccharide having several biomedical applications. This study simultaneously compares the anticancer activities of crude fucoidans from Fucus vesiculosus and Sargassum filipendula, and effects of low (LMW, 10-50kDa), medium (MMW, 50-100kDa) and high (HMW, >100kDa) molecular weight fractions of S. filipendula fucoidan against osteosarcoma cells. Glucose, fucose and acid levels were lower and sulphation was higher in F. vesiculosus crude fucoidan compared to S. filipendula crude fucoidan. MMW had highest the levels of sugars, acids and sulphation among molecular weight fractions. There was a dose dependent drop in focal adhesion formation and proliferation of cells for all fucoidan-types, but F. vesiculosus fucoidan and HMW had the strongest effects. G1-phase arrest was induced by F. vesiculosus fucoidan, MMW and HMW, however F. vesiculosus fucoidan treatment also caused accumulation in sub G1-phase. Mitochondrial damage occurred for all fucoidan-types, however F. vesiculosus fucoidan led to mitochondrial fragmentation. Annexin V/PI, TUNEL and cytochrome c staining confirmed stress induced apoptosis-like cell death for F. vesiculosus fucoidan but features of stress-induced necrosis-like cell death for S. filipendula fucoidans. There was also variation in penetrability of different fucoidans inside the cell. These differences in anti-cancer activity of fucoidans are applicable for osteosarcoma treatment.

IN VITRO ANTIOXIDANT AND CYTOTOXIC PROPERTIES OF FUCOIDAN FROM THREE INDIAN BROWN SEAWEEDS

Objective: In the present study, fucoidan extracted from three brown algae, Sargassum wightii, Turbinaria ornata, and Padina tetrastromatica, was purified, characterized, and evaluated for antioxidant and cytotoxic properties. Methods: Algal powders were sequentially extracted with five solvents based on polarity and residue was subjected to acidic extraction. The filtrates were precipitated for alginates, and resultant supernatant was precipitated for fucoidan. The precipitate was centrifuged; pellet dialyzed and lyophilized to yield crude fucoidan, which was purified by diethylaminoethyl cellulose chromatography and characterized by biochemical tests and Fourier-transform infrared (FT-IR) spectrometry. Solvent extracts and fucoidans were subjected to 2,2-diphenyl-1-picrylhydrazyl assay. Fucoidans were subjected to trypan blue cytotoxicity assay. Results: Antioxidant activity was highest in methanol extracts and Padina crude fucoidan, while lowest in hexane extracts and purified Sargassum fucoidan. Sargassum yielded the highest amount of fucoidan (7.14%). Total carbohydrates increased as Sargassum> Padina > Turbinaria, sulfates as Padina > Turbinaria > Sargassum, and protein content was 0.16±0.001%. Cytotoxicity increased in a dose-dependent manner; the highest and lowest for Padina at 200 mg mL-1 (40%) and 10 mg mL-1 (4%), respectively. Antioxidant and cytotoxic properties exhibited a positive correlation with sulfate content. FT-IR spectral values were characteristic to fucoidan. Conclusion: Fucoidans from the three algae effectively scavenged free radicals and showed good cytotoxic activity. There was a positive correlation between sulfate content and bioactivity of fucoidans, supporting its structure-function relationship. Thus, extracts and fucoidans from these algae are found to be potential candidates for pharmacological applications.

Perbandingan Profil Lipid pada Tikus Putih yang diberi Crude Fucoidan dari Rumput Laut Coklat (Sargassum polycystum) yang Diinduksi Diet Tinggi Lemak

Rumput laut coklat (Sargassum polycystum) merupakan salah satu tanaman yang menghasilkan aktivitas farmakologik yaitu sebagai anti aterosklerotik dengan kandungan kimia yang berpotensi adalah fucoidan. Penelitian ini bertujuan untuk mengetahui efektivitas crude fucoidan dalam menurunkan kadar kolesterol, LDL, TG, menaikkan kadar HDL dan Indeks Aterogenik. Pada tahapan optimasi digunakan 5 ekor hewan uji tikus untuk melihat pengaruh komposisi diet tinggi lemak yang diberikan terhadap profil lipid. Sedangkan pada tahap pengujian digunakan 30 ekor hewan uji yang dibagi menjadi 6 kelompok untuk melihat pengaruh diet tinggi lemak dan crude fucoidan yang diberikan terhadap profil lipid. Pemeriksaan profil lipid dilakukan sebanyak tiga kali yaitu pada hari ke-0, ke-14 dan hari ke-36 setelah itu dihitung indeks aterogeniknya. Pada hari ke-36 hewan uji di anestesi dan dinekropsi untuk selanjutnya dilakukan pengambilan darah melalui organ jantung. Hasil penelitian menunjukkan bahwa crude fucoidan dosis 200 mg dapat menurunkan kadar kolesterol total, LDL, TG, menaikkan kadar HDL dan memiliki penurunan indeks aterogenik serum tikus secara bermakna dibandingkan dengan dosis 100 mg dan 50 mg, tetapi tidak berbeda bermakna dengan kontrol positif atorvastatin dosis 10 mg.