Wps 3000 sampler

Batches (50 mL) of 1% (w/w) native and Bl-αAmy-treated potato starch were gelatinized according to the previously described method [38 (link)]. The samples were diluted to 0.2% (w/v) in 100 mM acetate buffer, pH 6, and filtered through 0.22 µm nylon filters (Frisenette, Knebel, Denmark). High-Performance Size-Exclusion Chromatography (HPSEC) was performed using an Ultimate iso-3100 SD pump with a WPS-3000 sampler (Thermo Scientific, Waltham, MA, USA) connected to an ERC RefractoMax 520 refractive index detector (Thermo Scientific, Waltham, MA, USA). The column employed was a Shodex SB-806 HQ column (300 × 8 mm) equipped with a Shodex SB-G guard column (50 mm × 6 mm) (Showa Denko K.K., Tokyo, Japan). The samples were run as previously described by [39 (link)] using external pullulan standards in the range of 342–805,000 Da (PSS Polymer Standards Service GmbH, Mainz, Germany). Molecular weight above the applied standard range was estimated based on the extrapolation of a third-degree polynomial expression.

The molecular weight distribution of the native and hydrolyzed fucoidan was estimated by high-performance size exclusion chromatography (HPSEC) using an Ultimate iso-3100 SD pump with a WPS-3000 sampler (Thermo Scientific, Waltham, MA, United States) connected to an ERC RefractoMax 520 refractive index detector (Thermo Scientific, Waltham, MA, United States). 100 μl of sample was loaded on a Shodex SB-806 HQ column (300 × 8 mm) equipped with a Shodex SB-G guard column (50 mm × 6 mm; Showa Denko K.K., Tokyo, Japan).
Elution was performed with 100 mM sodium acetate pH 6 at a flow rate of 0.5 ml/min at 40°C. External pullulan standards (PSS Polymer Standards Service GmbH, Mainz, Germany) were applied to establish a polynomial relationship between the logarithmic molecular weight and the corresponding retention times separating the polymer molecules in order to convert the retention times of the samples to molecular weights. Data visualization was performed in Python 3.8.

The molecular weights (MW) of fucoidan fractions were determined by High Performance Size Exclusion Chromatography (HP-SEC) using an Ultimate iso-3100SD pump with WPS-3000 sampler (Dionex, Sunnyvale, CA, USA) connected to an RI-101 refractive index detector (Shodex, Showa Denko K.K., Tokyo, Japan). The samples were dissolved in 1 ml HP-SEC buffer (100 mM sodium acetate, pH 6) and filtered through 0.22 μm filters. 100 µL of fucoidan samples (3 mg/mL) were injected into a Shodex SB-806 HQ GPC column (300 × 8 mm) coupled with a Shodex SB-G guard column (50 mm × 6 mm) (Showa Denko K.K., Tokyo, Japan) [56 (link)]. Elution was carried out at a flow rate of 0.5 mL/min at 40 °C [56 (link)]. Pullulan samples of molecular weight 1, 5, 12, 110, 400, and 800 kDa were used as standards [56 (link)].