Lc 10ad hplc pump

The TSNA MIP column was prepared following a modified method (Forzano et al., 2019 (link)) under slurry conditions using a Teledyne (Thousand Oaks, CA) packing pressure system. The end fitting and .2 μm frit from one end of a Restek (Bellefonte, PA) column assembly kit (50 mm × 2.1 mm ID, ¼” OD) was removed and attached to a reservoir with a glass frit (Figure 1). One Gram of TSNA MIP material from Biotage was suspended in 20 ml of a 1:1 (v/v) chloroform:ethanol solution to create a slurry, which was sonicated for 15 s. The mixture was poured into the reservoir and the system was capped and secured. Pushing solvent consisting of 1:1:1 (v/v/v) ethanol:isopropyl alcohol: toluene was used to push the slurry mixture through the column. Helium gas was used to degas the solvent during the packing process. The pushing solvent was slowly increased from .5 ml/min to 10 ml/min with a total system pressure around 2000 psi. The flow rate of the pushing solvent was kept at 10 ml/min for 1 hour before the system was disassembled and the previously removed frit and end fitting were reassembled on the column. The freshly packed column was washed with 100% acetonitrile at a flow rate of .5 ml/min for 60 min using an external Shimadzu LC-10AD HPLC pump (Kyoto, Japan). Three HPLC columns were packed with TSNA MIP material from the same lot.

The molecular weight and molecular weight distribution were determined by gel permeation chromatography (GPC). The chromatography was carried out at 35 °C temperature using HPLC system (Shimadzu Deutschland GmbH, Duisburg, Germany) fitted with LC-10AD HPLC pump and RID-6A refractive index detector coupled with a styragel HR4 column in an isocratic mode with the deaerated chloroform as mobile phase at a flow rate of 1 ml/min. The samples were prepared by dissolving 10 mg of polymer in 2 ml chloroform and filtered through 0.2 µm membrane filter (Millipore). 50 µl of the sample was injected and analyzed using “GPC for Class-Vp” software (Shimadzu Deutschland GmbH, Duisburg, Germany). The molecular weights were calculated from the elution volume of polystyrene standards with molecular weight range of 5610–275,000 Da (Polysciences Asia Pacific, Inc. Taiwan).