I. II. III. IV.

Phenolic Content of Marine Macroalga Industrial Applications Methods of Extraction Methods of Measurement a. Folin Ciocalteu Reagent i. Used in many studies ii. Results are to be reported as mol gallic acid equivalent (GAE)/ g wet tissue 1. (from http://www.medwelljournals.com/fulltext/?doi=jftech.2010.118.125) Evaluation
of antioxidant activity Determination of total phenolic content: The phenolic contents were determined using Follin-Ciocalteu reagent and expressed as Gallic Acid Equivalents (GAE) (Singleton et al., 1999).

2. The extracts were diluted with methanol, by taking 3 mL of methanol and 1 mL of crude
fruits extract solution.

3. To this sample solution, 1 mL of 5-fold diluted Folin-Ciocalteus reagent was added. 4. The contents were mixed well and kept for 5 min at room temperature followed by the
addition of 1 mL of 10% aqueous sodium carbonate.

5. After incubation at room temperature for one and half hour the absorbance of the
developed blue color was read at 760 nm (Shimadzu UV-1650 PC Shimadzu Corporation, Kyoto, Japan) against reagent blank.

6. Gallic acid (100-1000 mg mL-1) was used to construct the calibration curve. Results were
calculated as gallic acid equivalents (mg/100 g) of samples.

7. The reactions were conducted in triplicates and concentrations of phenolic compounds

were calculated according to the equation obtained from the standard dallic acid graph: A = 0.0044 (gallic g) + 0.6746 (R2: 0.9173). Folin part two (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3167909/)

b.

1. Total phenolics were determined according to the method mentioned by Prabhasankar et al. (2009). 2. In a test tube, 0.2 ml of the aqueous extract was mixed with 4.0 ml of 0.02 g/ml Na2CO3 and then 0.2 ml of 0.5 g/ml Folin Ciocalteaus phenol reagent was added. 3. The reaction mixture was mixed thoroughly and allowed to stand for 30 min at room temperature in the dark. 4. After incubation, the solution was measured at 720 nm against a blank. Results were expressed as mg gallic acid equivalents per 100 g dry weight (mg GAE/100 g DW).