Pullulan polysaccharide calibration kit

The molecular weight of H. erinaceus polysaccharide was determined by High-performance Gel Permeation Chromatography (HPGPC, 6000 high performance liquid chromatograph, Waters, MA, USA). A 300 mm × 7.8 mmid × 2 Ultrahydrogel™ Linear (Waters, Milford, MA, USA) was eluted with 0.1 M NaNO₃ (0.9 mL/min, 45 °C) and determined by a Waters 2410 RI detector. Calibrating the average molecular weight was performed using a pullulan polysaccharide calibration kit (Agilent Technologies, Santa Clara, CA, USA).

Fresh seedless chestnut rose (R. sterilis) fruits were collected in October 2021 on the farmland of Zhenning, Anshun, located in Guizhou Province, China (26°2′49.42″ N, 105°41′59.45″ E, 1247.6 m above sea level). Standard monosaccharides, 3-Phenylphenol, trifluoroacetic acid (TFA), 1,10-Phenanthroline, 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and 3-(2-Pyridyl)-5,6-diphenyl-1,2,4-triazine-p,p′-disulfonic acid monosodium salt hydrate were purchased from TOKYO Chemical Industry Co., Ltd. (Tokyo, Japan). Sodium hydroxide was purchased from Thermo Fisher Co., Waltham, MA, USA. The pullulan polysaccharide calibration kit was purchased from Agilent Co., Santa Clara, CA, USA. α-Glucosidase, p-nitrophenyl-α-d-glucopyranoside (PNPG), and serum albumin (BSA) were obtained from Yuanye Bio-technology (Shanghai, China). All additional solvents and chemical reagents were acquired locally and were of either HPLC or analytical quality.

The average molecular weight of fucoidan from Ecklonia cava was determined by gel permeation chromatography (GPC) using a Shodex SB 804 HQ OHPak column (30 cm × 8 cm) equilibrated with phosphate buffer (10 mM, pH 7) for 60 min. A pullulan polysaccharide calibration kit (Agilent Technologies, Santa Clara, CA, USA) was purchased and used as a molecular weight standard.