HPLC Will Survive and Thrive
Those who thought HPLC had peaked in the 1990s were sorely mistaken – but can the technique continue to adapt to increasing demands?
High-performance liquid chromatography (HPLC) was born half a century ago, and has since become one of the most widely used analytical techniques in the world. The demand for higher separation efficiency, faster analyses and greater throughput has been the driving force for extraordinary advances in stationary phases. One of the secrets of HPLC’s success is that column technology caters for all tastes, and it is capable of continuous renewal.
The versatility of stationary-phase chemistry, or surface modification, allows for the design of efficient and selective packing materials for the separation of nonpolar, polar, ionizable, or chiral analytes, small molecules or large biomolecules. Particle sizes have decreased from 50 µm to less than 2 µm. Core-shell particles have become the favorite packing materials for many users, and not only for the separation of macromolecules (even though those particles were originally developed for the efficient separation of biomolecules). Silica-based monolithic columns have become a popular alternative for efficient separations at moderate pressure drops. We are going to see many more exciting new developments in the field of monoliths, as the short columns developed for fast HPLC can be operated at higher pressures than the conventional silica-based monoliths.
It has become clear that column hardware has an essential impact on band broadening, and thus on separation efficiency. In a typical 50×2.1 mm column developed for fast liquid chromatography, the band broadening of an unretained analyte in the 50 mm-long chromatographic bed is about the same as in the two 1 mm-long frits (1). To produce higher-efficiency columns for fast HPLC, the optimization of frit technology will be of utmost importance. Furthermore, instrumentation must develop so that the integration of the column in the instrument will not introduce dead volumes at the connecting elements, so that extra-column band broadening is kept to a minimum.
In the 1990s, many chromatographers thought that HPLC had matured and such a level that further development was possible only in incremental steps. But, soon after that, we witnessed the introduction of ultra-performance liquid chromatography, sub-2 µm and core-shell particles.
Now, we wait impatiently to find out how 3D-printing technology will impact on HPLC column technology. After the success of comprehensive multidimensional chromatography, we are anxious to see what spatial two-dimensional and three-dimensional HPLC will bring for us.
The perpetual development of column technology guarantees that HPLC will remain a success story. Some of the ideas being suggested for future developments may seem utopist, but given the huge and unforeseen developments seen recently in stationary-phase technology, we would be wise to keep our minds open.
For more discussion on HPLC’s place in modern analytical science click here. To hear from HPLC 2017 Prague chairs, Michal Holčapek and František Foret, visit: tas.txp.to/0117/HPLC2017
HPLC 2017 Prague takes place
June 18–22 at Prague Conference Centre.
www.hplc2017-prague.org
- N Lambert et al., “Comparison of the kinetic performance of different columns for fast liquid chromatography, emphasizing the contributions of column end structure”, J Chromatogr A, 1473, 99–108 (2016).
Attila Felinger is Professor at the University of Pécs, Pécs, Hungary.
