Au 950 autosampler

SEC was performed out on an HLC-8120 system (Tosoh, Tokyo, Japan) equipped with a combination of TSKgel AW-4000 and AW-2500 columns (150 mm × 6 mm ID) (Tosoh), eluted with dimethylsulfoxide (DMSO) containing 10 mM LiBr at a flow rate of 0.15 mL/min at 65 °C. The polydispersity index (M_w/M_n) was calculated from a calibration curve of standard PEGs (Agilent Technologies, Wilmington, DE, USA). For SEC with fluorescence detection, the measurements were performed on a Gulliver system (Jasco, Tokyo, Japan) consisting of a DG-2080–53 degasser (Jasco), a PU-980 pump (Jasco), an AU-950 autosampler (Jasco), a CO-965 column oven (Jasco), an FP-920 fluorescence detector (excitation wavelength: 488 nm, emission wavelength: 515 nm) (Jasco), an RI-2031 Plus refractive index detector (Jasco), and a combination of TSKgel α-4000 and TSKgel α-2500 columns (300 mm × 7.8 mm ID) (Tosoh). The solutions (50 μL) were injected into the SEC system, and the system was eluted with a mixture of water and DMSO (volume ratio 50:50) at a flow rate of 0.3 mL/min at 40 °C. ¹H NMR spectra were recorded on a Bruker Avance III 500 MHz spectrometer (Bruker BioSpin, Rheinstetten, Germany). FTIR spectra were recorded on a Spectrum 100 FTIR spectrometer (Perkin Elmer, Wellesley, MA, USA). The sample powder was mixed with KBr and pellets were prepared for FTIR measurements.

¹H nuclear magnetic resonance (NMR) spectra were recorded in D₂O at 25 °C using a Bruker Avance III 400 MHz spectrometer (Bruker BioSpin, Rheinstetten, Germany). Fourier transform infrared (FT-IR) spectra were recorded on a Spectrum 100 FT-IR spectrometer (Perkin Elmer, Wellesley, MA, USA). The samples were ground to powder form, which was blended with KBr to prepare pellets for measurements. Aqueous-phase size exclusion chromatography (SEC) measurements were performed using a high-performance liquid chromatography setup consisting of an AU-950 autosampler (Jasco, Tokyo, Japan), PU-980 pump (Jasco, Tokyo, Japan), DG-2080-53 degasser (Jasco, Tokyo, Japan), CO-965 column oven (Jasco, Tokyo, Japan), RI-2031 Plus refractive index detector (Jasco, Tokyo, Japan), and a combination of TSKgel G4000PW_XL and G2500PW_XL columns (300 mm × 7.8 mm ID; Tosoh, Tokyo, Japan). The sample solutions (10 mg mL⁻¹, 50 μm) were injected into the system and eluted with a 100 × 10⁻³ M NaNO₃ solution at a flow rate of 0.4 mL min⁻¹ at 40 °C [30 (link)].