The new Hamilton PRP-C18 HPLC column allows efficient separation processes under extremes of pH, high pressure conditions and temperatures of more than 100˚C. This simplifies the method development in reversed-phase chromatography.
Reversed-phase chromatography is the most frequently utilised technique for HPLC separations. The pH of the mobile phase plays a decisive role in the efficiency of separations. However, the low resistance of conventional HPLC columns still sets a limit to this advantage because most columns cannot handle pH values above 7 and below 3 without suffering a loss in performance. Hamilton has now developed a PRP-C18 column that can be used with virtually any mobile phase at any pH.
The need for pH tolerant HPLC columns becomes more obvious in drug discovery as more than 70% of all drug compounds are amines that can only be separated at higher pH values above 7. With the Hamilton PRP-C18, neutral amines or other organic bases can be isolated under alkaline conditions in a highly efficient manner without any degradation of the column. Nine structurally-diverse, strongly basic drug compounds were resolved in less than two minutes using a mobile phase with a pH of 11.5 and a fast acetonitrile gradient at 80˚C.
One reason for the prolonged efficiency is the way the column can be cleaned. Unlike traditional ODS C18 columns, the PRP-C18 can be regenerated using strong acidic and basic solutions without causing damage. Contaminants that are normally bound irreversibly to ODS columns and may negatively affect analytical results can be easily washed away with one molar sodium hydroxide or sulfuric acid. The result is sustained efficiency, stable retention times and a high-degree of reproducibility.
The PRP-C18 is also completely inert towards other problematic conditions in reversed-phase chromatography: high pressures and temperatures. The chemical and mechanical stability of the column allows operation at up to 5000 psi (350 bar) and more than 100˚C. Analyses at high temperatures reduces run times, cutting down on organic solvent consumption. The highly cross-linked PRP-C18 resin packing does not swell or shrink under extreme operating conditions, providing the flexibility to explore other conditions in methods developments.
The trend in modern chromatography is towards smaller column inner diameters using smaller particles, which can be used under extreme conditions in order to increase laboratory productivity. The Hamilton PRP-C18 takes advantage of this trend for polymer resins and thus combines extreme column stability with excellent separating performance.
Hamilton Bonaduz AG, +41 81 660 67 41, mnipius@hamilton.ch, www.hamilton.ch.