*4.4. Chemical Analysis*

The proximate composition of the chemical constituents in the samples was determined according to the methods described in AOAC 2002 [27]. The adopted AOAC methods for the evaluation of ash, protein, lipid, and carbohydrates were dry ashing in a furnace at 600 ◦C for 5 h, Kjeldahl digestion, Soxhlet extraction, and phenol-sulfuric assay. The polysaccharide and sulfate content of the purified fractions were respectively analyzed using the standard phenol-sulfuric assay and BaSO4 precipitation methods with some minor modifications [28]. Additionally, the protein and polyphenol content were estimated using the commercial BCA protein assay kit and by Folin–Ciocalteu method, respectively [29]. Monosaccharide composition was analyzed by a CarboPac PA1 cartridge column connected to an ED50Dionex electrochemical detector after hydrolysis by trifluoroacetic acid [1].
