The limits of the separation power of unidimensional column liquid chromatography.

The practical limit of the separation power of HPLC depends on time, money, and skill. That is it depends on the time available for the analysis, on the quality and performance of the pump and hardware and particularly on the maximum pressure at which the pump can deliver the mobile phase to the column, and on the temperature at which the column can be operated. It also depends on the properties of the packing material selected (e.g., its particle size, its pore geometry, and its connectivity) and on the packing method used since it affects the coefficients of the HETP equation. Finally, it depends on the thermal stability of the sample and the packing material. The complexity of the sample also plays an important role in that it determines whether the analysis should be made under isocratic, isothermal conditions, in gradient elution, in temperature programming, or with a combination of both types of programming. The various phenomena that affect column properties and separation performance are discussed. Past achievements suggest that columns providing efficiencies in excess of a million plates in less than 1 day are within the grasp of current technology. The possibility of further advances are considered.