Titanium-scaffolded organic-monolithic stationary phases for ultra-high-pressure liquid chromatography

• Highly robust organic-monolithic columns were prepared in a titanium scaffold.
• Columns were prepared repeatedly and the metal scaffold could be recycled easily.
• Columns were sustainably used for separations at ultra-high pressures.
• Columns were successfully used with gradients starting from 100% acidified water.
• Reasonable efficiencies were obtained for separating proteins.

Organic-polymer monoliths with overall dimensions larger than one millimetre are prone to rupture – either within the monolith itself or between the monoliths and the containing wall – due to the inevitable shrinkage accompanying the formation of a cross-linked polymeric network. This problem has been addressed by creating titanium-scaffolded poly(styrene-co-divinylbenzene) (S-co-DVB) monoliths. Titanium-scaffolded monoliths were successfully used in liquid chromatography at very high pressures (up to 80 MPa) and using gradients spanning the full range of water–acetonitrile compositions (0 to 100%). The kinetic-performance of (50-mm long) titanium-scaffolded monoliths was compared to that of similar monolith created in 1-mm i.d. glass-lined tubing at pressures up to 50 MPa. The peak capacities obtained with the titanium-scaffolded column was about 30% lower. An increased Eddy-diffusion, due to the pillar-structure, and a decreased permeability are thought to be the main reasons for this reduced kinetic-performance. No decrease in performance was observed when the titanium-scaffolded columns were operated at pressures of 80 MPa for up to 12 h. The column-to-column repeatability (n = 5) was acceptable in terms of observed peak widths at half heights (RSD ca. 10%) The run-to-run repeatability (n = 135) in terms of retention times and peak widths at half height were found to be good. Titanium-scaffolded columns coupled in series up to a combined length of (200 mm) were used for the analyses of a complex Escherichia coli protein sample. Our experiments demonstrate that columns based on titanium-scaffolded organic-polymer monolith can be operated under strenuous conditions without loss in performance. The titanium-scaffolded approach makes it feasible to create organic-polymer monoliths in wide-bore columns with accurate temperature control.