Cbm 20a

Dried ANE samples were placed inside a furnace at 600°C for 6 h in order to obtain and quantify the ash content. Total sugars were quantified according to Rioux et al. (2007) (link). Total polyphenol content was determined spectrophotometrically following the method of Goñi et al. (2018) (link). The content of unidentified organic components was calculated by difference to the total organic amount. The molecular weight (M_w) distribution of carbohydrates from different samples was analyzed using high performance size exclusion chromatography-refraction index detector (HPSEC-RID). The HPSEC Shimadzu system consisted of a system controller CBM-20A, a solvent delivery module LC-20 AD, an online degasser DGU-20A5, an autosampler SIL-20ACHT, a refraction index detector (Varian Prostar 350 RID), and an LC workstation. HPSEC analysis was performed using 4 PL aquagel-OH MIXED-H columns in tandem (8 μm, 300 × 7.5 mm; Agilent). The mobile phase (0.1 M NaAc/0.1 M Na₂SO₄ buffer pH 7.8) was used as isocratic elution at room temperature. The flow rate and injection volume were set to 1 ml min⁻¹ and 40 μl, respectively. M_w values were calculated from the measured retention times through a calibration curve made with dextran standards.

The molecular weight (M_w) distribution of carbohydrates of the four brown seaweed extracts was detected and measured using high-performance size exclusion chromatography with a refraction index detector (HPSEC-RID). The HPSEC Shimadzu system consisted of a system controller CBM-20A, a solvent delivery module LC-20AD, an online degasser DGU-20A5, an autosampler SIL-20ACHT, a refraction index detector (Varian Prostar 350 RID) and an LC workstation. HPSEC analysis was performed using PL aquagel-OH MIXED-H columns (8 μm, 300 × 7⋅5 mm; Agilent). The mobile phase (0⋅1 M NaAc/0⋅1 M Na₂SO₄ buffer, pH 7⋅8) was used as the isocratic elution at room temperature. The flow rate and injection volume were set to 1 ml/min and 40 μl, respectively. A molecular weight calibration curve was constructed with the retention time values of known dextran standards (Sigma-Aldrich, MO, USA). For the analysis of the extracts, the measurable range was divided into four segments (>100, 50–100, 10–50 and <10 kDa). An average M_w for each extract within each range was determined, and relative peak area values were calculated using the LCsolution software (Shimadzu, Ireland).