Gel-Permeation Chromatography (GPC)

GPC with THF as mobile phase was conducted using a Shimadzu system fitted with a Wyatt DAWN DSP multi-angle laser light scattering detector (690 nm, 30 mW) and a Wyatt OPTILAB EOS interferometric refractometer (690 nm). Three Agilent PLgel columns (MIXED-C; 5 μm bead size) were employed, operating at 1 mL per minute at a column temperature of 45 °C. To process the GPC data, the program ‘Astra’ by Wyatt technologies was used. All samples were filtered through 0.45 μm nylon filters prior to injection. When aqueous-phase GPC was employed, a separate Shimadzu liquid chromatography system was utilized, equipped with a Shimadzu RID-10 refractometer (λ = 633 nm), using three Waters Ultrahydrogel columns in series ((i) 250 Å porosity, 6 μm diameter bead size; (ii) and (iii) linear, 10 μm diameter bead size), operating at room temperature. The eluent was Milli-Q water containing 20% v/v acetonitrile and 0.1% w/v TFA at a flow rate of 0.5 mL·min⁻¹. The molecular weight of the analyte was determined using a calibration based on narrow molecular weight distribution poly(ethylene glycol) standards.