Chromatography
Gel Permeation Chromatography is performed on a Tosoh EcoSEC HLC-8320GPC System equipped with a series of TSKgel SuperMultiporeHZ-H, -M, and -N columns. The instrument is equipped with an RI and UV absorbance detector (UV-8320). The instrument is run with HPLC-grade tetrahydrofuran as the eluent and toluene as an internal standard, at a flow-rate of 0.500 mL/min, temperature of 40 °C, and injection volume of 50-100 μL. Prior to injection, all samples are fully dissolved in the mobile phase at a concentration of ~1-5 mg/mL and passed through a 0.45 μm syringe filter. Molecular weight and molecular weight distributions for the samples are estimated using a calibration curve generated from a set of poly(styrene) (Tosoh Biosciences) or poly(methyl methacrylate) (Agilent) standards. This instrument is also equipped with a LenS3 MALS-V(multi-angle light scattering-viscometer) dual detector, using a 505 nm laser to directly measure absolute molecular weight (Mw), radius of gyration (Rg) and intrinsic viscosity (IV), including macromolecules below 10 nm in gyration size (Rg).
Gel Permeation Chromatography (GPC)
The Biotage Selekt Flash Chromatography System (SEL-2SW Biotage® Selekt) can be utilized for both normal and reverse phase column chromatography. This system contains two channels which enables the user to run two separations on one instrument. It has a single collection bed with room for three test-tube racks. A UV-VIS Detector (198–810 nm, amplitude range: 0 to 6400 mAU (Absorbance Units);accuracy of ±5 mAU) which enables the user to collect sample only when something is detected and can also be set to collect only within a certain wavelength range. It has a two-flow rate range from 1-300mL/min. in 1 mL/min increments with a pressure range of 0-30 bar. The current solvents we have available are: HPLC Grade hexanes, ethyl acetate, acetonitrile, water, dichloromethane, and methanol. The system can hold and run up to 4 solvents at the same time. The software for the instrument is SEL-SPK Biotage® Selekt, Spektra Software Package Accessories and we utilize the pre-packed Biotage Sfär Columns (FSRD-0445-0XXX Silica Duo - Samplet® SupportSilica D) (FSUD-0443-0XXX Silica Duo -Samplet® Support Silica HC D) (FSUD-0401-0XXX) Silica C18 Duo - Samplet® Support C18 D)
Automated Flash Chromatography
The LC diagrams were recorded by the Agilent 1260 Infinity II Binary LC System, which is equipped with a G7165AMultiple Wavelength Detector. The detector recorded UV-Vis spectrum ranging from 190 - 950 nm with the fast repetition rate of 120 Hz. The system is equipped with a C18 reversed phase column (InfinityLab Poroshell 120) as the stationary phase. The mobile phase is HPLC grade water and acetonitrile, and the typical flow rate is 1 mL/min at room temperature. The injection volume is 1 to 100 μL.
