Through-pore polymerization in polar high-performance liquid chromatography columns allowing scanning electron microscopy based imaging of the packing order

• A new in-column methacrylate-based polymerization procedure was developed for imaging the internal morphology of bare silica HPLC columns Polymerization performed in a commercial and home-made column The obtained silica/polymeric rod can be mechanically cut allowing cross section SEM allows mapping local external porosity packed could assistance to steer optimization packing processes columns. To allow an enhanced understanding order columns, this work methodology immobilizing native polar particles is based on methyl methacrylate (MMA) ethylene glycol dimethacrylate (EGDMA) as cross-linker interstitial pores Subsequent mechanical cutting then scanning electron microscopy (SEM) imagery cross-sections bed. In way, efficiency with 4.6 mm inner diameter 150 length comprising same material 5 µm are compared. used e.g. hydrophilic interaction liquid chromatography (HILIC) normal phase LC. confirm feasibility methodology, conventional methods evaluation column, porosity, also employed. information compared showed trend measurements via inverse size exclusion approach, illustrating its potential application study micro-heterogeneity guide process