Seaweed health benefits determination

Seaweed has a wide array of applications in food, fertilizers, phycocolloids, and cosmetic ingredients for nutraceuticals and pharmaceuticals. It is, however, considered underutilized as a resource.

• Great nutrient source of ucoidan, phenolics, fatty acids, polysaccharides, vitamins, sterols, tocopherols, terpenoids, and phycocyanins.
• UV protective functions
• Powerful antioxidant  
• Acts as a sustainable feedstock for biogas production
• Photosynthetic pigments have functional food applications and display health benefits in various ways. These are credited to its anticancer, anti-inflammatory, antidiabetic, anti-obesity, antiangiogenic, neuroprotective and other biological activities

The researchers have identified a key issue in the underutilization of seaweed. It is said that not enough work has been done to fully explore its application capacities.

...further work needs to be performed to identify the metabolites components of seaweed that are useful for industrial and pharmaceutical applications.

It is believed that a more holistic method is needed to confirm health benefits of seaweed through biochemical profiling.

• Evaluate chemical compositions
• Research α-glucosidase inhibitory and cytotoxic activities of extracts (types of seaweed used are U. intestinalis, H. macroloba, and S. ilicifolium) against five cancer cell lines including MCF-7 cells, MDA-MB-231 cells, HT-29 cells, Hep G2 cells, and 3T3 cells
• Investigate for the commercial cultivation of edible seaweed as human food

A portion was checked for taxonomic identification. Afterwards, the seaweed is once more washed, frozen overnight, lyophilized in a freeze dryer until constant biomass weight is achieved. Then, these freeze-dried samples are pulverized and sieved.

Finely prepared sample powder is mixed in methanol; this resulting solution is sonicated for an extraction. The solvent extract is filtered, and dried residue undergoes methanol mixing once more. The process repeats a third time. The extracts are dried in a rotary evaporator.

The newly dried lipid samples are stored. At this point, the total carotenoids and chlorophyll a and b contents of dried seaweed lipid extracts is measured. For exact figures, read the published study.

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α-Glucosidase Inhibition Assay

The researchers, at this point, determine the assay for a-glucosidase inhibition activity. ρ-Nitrophenyl-ρ-D-glucopyranoside (PNPG) substrate is used. Seaweed extracts are prepared, mixed in a 96-well microplate and incubated at room temperature for 5 minutes. 75 μL of PNPG is added to wells consisting of sample, blank substrate, and negative/positive controls. Leftover wells are filled with 75 μL of 30 mM phosphate buffer. The wells are incubated for 15 minutes at room temperature. Reaction mixtures are applied with stopping agent. Absorbance is measured with a spectrophotometer set to 405nm. The resulting a-glucosidase inhibition activity is expressed as percentage (%) of inhibition.

Cells and culture conditions

The following are used:

• MCF-7 cells (human breast adenocarcinoma cell line, estrogen receptor positive)
• MDA-MB-231 cells (human breast adenocarcinoma cell line, estrogen receptor negative)
• HT-29 cells (human colorectal adenocarcinoma cell line)
• Hep G2 cells (human liver hepatocellular carcinoma cell line)
• 3T3 cells (mouse embryonic fibroblast cells)

As for the condition, these cells were maintained for research purposes in RPMI medium which is supplemented with 10% fetal calf serum (FCS), 100 unit/ml penicillin, and 0.1 mg/ml streptomycin.

MTT Assay Conditions

The sample extracts are tested for cytotoxicity levels. The extracts are tested on the above-mentioned cells. Protocol used in the anticancerous Effect of Typhonium flagelliforme on Human T4-Lymphoblastoid Cell Line CEM-ss research is used to determine the cytotoxicity using MTT assay. At the beginning, the cell culture was prepared (at a concentration of 1 × 105 cells/mL) and plated (100 μL/well) into 96-well plates. Then incubation takes place, diluted ranges are dissolved in dimethyl sulfoxide (DMSO) and concentration of the DMSO is 0.1% (v/v).

Concentrations are plated as triplicate with untreated cell controls and blank cell-free control in each. A positive control was maintained as well. 68-hour incubation period followed, 20 μL of MTT (5 mg/mL) was added to individual wells to allow for formazan crystals formation and afterwards incubation took place to then remove the media later.

Diligent notes are made; the absorbance is recorded, percentage of cellular viability is noted, concentration which inhibited 50% of the cellular growth (IC50 value) is measured, inhibitory rate of cell proliferation is calculated, cytotoxicity of sample on cancer cells was expressed as IC₅₀ values, and further, mycoplasma contamination tests on these samples resulted as negative.

Gas Chromatography-Mass Spectrometry Analysis

This part of the experimentation is carried out to characterize and quantify bioactive substances in the seaweed extracts. The study made use, with modifications, of course, of the method found in research surrounding the triterpene Glycosides from Sea Cucumber Holothuria leucospilota.

This study on U. intestinalis, H. macroloba, and S. ilicifolium seaweed have shown:

• They can be good sources of bioactive, nutritionally and physiologically essential compounds
• These compounds have opportunities for applications in nutraceuticals and pharmaceutical industries in the form of food and medicine
• Seaweed pigments (total carotenoids, chlorophylls a and b) posses antidiabetic and anticancer potential

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