Gpc software

The weight-average molecular weight (Mw), number-average molecular weight (Mn), and polydispersity index (Mw/Mn) of RSPs were measured by high-performance gel permeation chromatography (HPGPC, Agilent 1260, Agilent Co., Santa Clara, CA, USA) coupled to a refractive index detector (RID, Agilent) at 35 °C. PL aquagel-OH 60 gel column (25 mm × 300 mm, 8 μm, Agilent, Cheadle, UK) and PL aquagel-OH 50 gel column (25 mm × 300 mm, 8 μm, Agilent, UK) were used as columns, and the mobile phase was KH₂PO₄ (0.02 M, pH 6.0) with a flow rate of 0.6 mL/min. The molecular weight distribution of the sample was calculated using the GPC software (version A.02.02, Agilent Co., Waldbronn, Germany), and the pullulan polysaccharide standards (molecular mass, 667, 6300, 22,000, 49,700, 216,000, 334,000, and 739,000 Da) were used as molecular mass markers.

The molecular weight of EPS was analyzed by gel permeation chromatography (GPC) (20 (link)). EPS sample (15 mg) dissolved in 1 mL ultrapure water was filtered through a 0.22-μm filter and put into a 5-mL sample bottle, and then ultrapure water containing NaNO₃ (0.2 M) and NaH₂PO₄ (0.01 M) was added to the scale line and ultrasonicated. The ultrasonic dispersion was used for molecular weight determination. GPC system (Waters 1525 Isocratic HPLC Pump) was equipped with a PL aquagel-OH MIXED 8 μm chromatographic column and a differential detector (Waters 2414 Refractive Index Detector). Ultrapure water containing NaNO₃ (0.2 M) and NaH₂PO₄ (0.01 M) was used as the mobile phase, the flow rate was 1 mL min^–1, and the column temperature was 30°C. The standard sample was narrow distribution polyethylene glycol provided by Polymer Standards Service-USA, Inc., with molecular weights (Mp) of 330,000, 176,000, 82,500, 44,000, 25,300, 20,600, 12,600, 7130, 4290, 1400, 633, and 430, respectively, and the data were analyzed using Agilent GPC software.

High-performance gel permeation chromatography (HPGPC) was utilized for the determination of RSP’s molecular weights with 3 mg of each RSP sample used. Separation utilized two Agilent PL aquagel-OH gel columns (60 and 50 series, 25 mm × 300 mm, 8 μm) with KH₂PO₄ (0.02 M, pH 6.0) at 0.6 mL/min, 30 °C, and a 20 μL injection volume. Calibration involved seven pullulan polysaccharide standards (667 Da to 739,000 Da). GPC software (version A.02.02, Agilent Co., Waldbronn, Germany) was used to determine the molecular weight distribution, including the number-average (Mn) and weight-average (Mw) molecular weights and the polydispersity index (Mw/Mn).