Optimized Extraction,Preliminary Characterization and Evaluation of the in Vitro Anticancer Activity of Phlorotannin-Rich Fraction from the Brown Seaweed,Cystoseira sedoides

Brown seaweeds produce useful bioactive substances with high cosmetic and pharmacological values due to the presence of antioxidant derivatives, mainly phlorotannins (PHT), which are of particular interest. A central composite rotatable design (CCRD) with three variables (extraction time, dry material-to-solvent ratio and ethanol concentration) and two responses was performed to optimize the microwave-assisted extraction (MAE) of phlorotannins from Cystoseira sedoides using response surface methodology (RSM). Results showed that the independent variables significantly affected both phlorotannin content and the scavenging capacity. The optimum operating conditions were extraction time, 101.74 sec; dry material-to-solvent ratio, 1:10 g/mL; and ethanol extraction, 50%. Under these conditions, the predicted values of PHT content and radical scavenging activity-IC50 were close to the observed values and were 383.887 µg PGE/g Dm and 18.353 µg/mL, respectively. Characterization of the phlorotannin-rich fraction was conducted by high-performance liquid chromatography and Fourier transform infrared spectroscopy. The evaluation of the anticancer activity against MCF-7 breast cancer cell line showed a potent activity to trigger apoptotic death in more than a half of the MCF-7 cells, with an estimated IC50 value of 78 μg/mL. In addition, this fraction induced a notable growth regression effect on 3D spheroids model in a concentration-dependent manner, with a growth rate of about 1.17, at 200 µg/mL.

Abbreviations: CCRD: Central composite rotatable design; Dm: Dry material; DMBA: 2,4-dimethoxy benzaldehyde; DMSO: Dimethyl sulfoxide; DPPH: 1,1-diphenyl-2-picrylhydrazyl; EDTA: Ethylene diamine tetraacetic acid; FBS: Fetal bovine serum; FT-IR: Fourier transform infrared; HPLC: High-performance liquid chromatography; IC₅₀: Half-maximal inhibitory concentration; MAE: Microwave-assisted extraction; Min: minutes; PBS: Phosphate buffered saline; PGE: Phloroglucinol equivalent; PHT: Phlorotannins; PHT-SED: phlorotannins derived from Cystoseira sedoides; PI: Propidium iodide; RSA: Radical scavenging activity; RSM: Response surface methodology; Sec: seconds     

The role of seaweed as a potential dietary supplementation for enteric methane mitigation in ruminants: Challenges and opportunities

Seaweeds are macroalgae, which can be of many different morphologies, sizes, colors, and chemical profiles. They include brown, red, and green seaweeds. Brown seaweeds have been more investigated and exploited in comparison to other seaweed types for their use in animal feeding studies due to their large sizes and ease of harvesting. Recent in vitro and in vivo studies suggest that plant secondary compound-containing seaweeds (e.g., halogenated compounds, phlorotannins, etc.) have the potential to mitigate enteric methane (CH₄) emissions from ruminants when added to the diets of beef and dairy cattle. Red seaweeds including Asparagopsis spp. are rich in crude protein and halogenated compounds compared to brown and green seaweeds. When halogenated-containing red seaweeds are used as the active ingredient in ruminant diets, bromoform concentration can be used as an indicator of anti-methanogenic properties. Phlorotannin-containing brown seaweed has also the potential to decrease CH₄ production. However, numerous studies examined the possible anti-methanogenic effects of marine seaweeds with inconsistent results. This work reviews existing data associated with seaweeds and in vitro and in vivo rumen fermentation, animal performance, and enteric CH₄ emissions in ruminants. Increased understanding of the seaweed supplementation related to rumen fermentation and its effect on animal performance and CH₄ emissions in ruminants may lead to novel strategies aimed at reducing greenhouse gas emissions while improving animal productivity.     

鼠尾藻多酚分级组分的抑菌活性研究

采用平板生长抑制法对鼠尾藻Sargassum thunbergiikuntze多酚化合物各分级组分进行抑菌活性研究。结果表明,在一定浓度范围内各分级组分对副溶血弧菌、哈维氏弧菌、沙蚕弧菌、金黄色葡萄球菌、大肠杆菌、鳗弧菌、溶藻弧菌和四联微球菌等受试菌株均有抑菌活性。其中,分级组分I(Mr5.0×103)的抑菌效果优于其他组分,对副溶血弧菌、哈维氏弧菌和沙蚕弧菌的最低抑菌浓度(MIC)值为900μg/ml,对金黄色葡萄球菌、大肠杆菌、鳗弧菌和溶藻弧菌的MIC值为1 800μg/ml,对四联微球菌的MIC值为3 600μg/ml。另外,组分I(Mr5.0×103)显示出较好的热稳定性,在pH 3~4时对受试菌的抑菌活性最强。     

Extraction Optimization for Phlorotannin Recovery from the Edible Brown Seaweed Ecklonia Cava

Phlorotannins, marine polyphenols from the brown seaweeds, are a promising biologically active substance. This study aimed to optimize the extraction conditions for phlorotannin recovery from the brown seaweed Ecklonia cava using response surface methodology. Ethanol concentration (X₁, %), extraction temperature (X₂, °C), and extraction time (X₃, h) were chosen as independent variables. The dependent variables were total phlorotannin content (TPhC; Y₁) and yield of phlorotannins (Y₂). Estimated optimal conditions were as follows: TPhC (Y₁): X₁ = 95%, X₂ = 80°C, X₃ = 22.8 h, and yield of phlorotannins (Y₂): X₁ = 88.3%, X₂ = 80°C, X₃ = 24 h. Predicted values of dependent variables under each optimal condition were TPhC (Y₁) = 570.2 mg phloroglucinol equivalents/g (mg PGE/g, dry basis) and yield of phlorotannins (Y₂) = 7.8% and were similar to the experimental values. In addition, content of dieckol, which is known as an indicator of phlorotannin products, was measured. Dieckol contents of the phlorotannin extracts with the highest (565.7 mg PGE/g) and lowest (315.4 mg PGE/g) TPhC were 49.1 and 34.4 mg/g (dry basis), respectively.     

Identification of Polyphenols from Chilean Brown Seaweeds Extracts by LC-DAD-ESI-MS/MS

Polyphenol profiles were characterized in extracts of three Chilean brown seaweeds, Durvillaea antarctica (Chamisso) Hariot, Lessonia spicata (Suhr) Santelices, and Macrocystis integrifolia (Bory) by liquid chromatography with mass spectrometry detection (LC-MS/MS). Phlorotannins with different degrees of polymerization were identified in D. antarctica (trimers to octamers) and L. spicata (trimers to tetramers). No signals related to phlorotannins compounds were detected in M. integrifolia. L. spicata and M. integrifolia showed a great variety of flavonoid compounds in comparison with D. antarctica, mainly identified as glycoside forms in all the extracts. The antioxidant activity of brown seaweed extracts measured by ferric reducing power (FRAP) and oxygen radical absorbance capacity (ORAC) was significantly higher in D. antarctica, followed by M. integrifolia and L. spicata, in line with the total phenolic (TP) content. However, D. antarctica and M. integrifolia showed similar activity for free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) in spite of the differences found in TP content. D. antarctica as well as L. spicata would represent a potential source of phlorotannins, whereas M. integrifolia could be considered as an alternative source of flavonoids. The identification of polyphenols in extracts of Chilean brown seaweeds opens innovative opportunities for their use in the food and pharmaceutical industries.