Fast separation of biological macromolecules on non-porous, microparticulate columns.

The use of high-performance liquid chromatographic columns for the separation of proteins and nucleic acids is gradually increasing in biochemical laboratories. The efficiency of these columns for such separations has been much lower than that achievable for the separation of smaller molecules. Non-porous microparticulate packings are the logical answer one arrives at after consideration of the chromatographic behaviour of proteins. Non-porous stationary phases are described for the separation of proteins, peptides and nucleic acids. The stationary phases used are TSK-Gel NPR-C18, TSK-Gel NPR-DEAE, TSK-Gel NPR-SP and HYTACH MicroPell C18. A number of fundamental properties of columns based on these sorbents were evaluated, such as permeability, retention behaviour towards small and large molecules, load capacity and stability. Instrumental requirements for these columns are discussed and some applications described.