Perspectives on Recent Advances in the Speed of High-Performance Liquid Chromatography

Abstract

Perhaps the most consistent trend in the development of HPLC since its inception in the 1960’s has been the continuing reach for ever faster analyses. The pioneering work of Knox, Horvath, Halasz, and Guiochon set forth a theoretical framework that was used early on to improve the speed of HPLC, primarily through the commercialization of smaller and smaller particles. Over the past decade, approaches to improving the speed of HPLC have become more diverse, and now practitioners of HPLC are faced with the difficult task of deciding which of these approaches will lead them to the fastest analysis for their application. Digesting the rich literature on the optimization of HPLC is a difficult task in itself, which is further complicated by contradictory marketing messages from competing commercial outlets for HPLC technology. In this perspectives article we provide an overview of the theoretical and practical aspects of the principal modern approaches to improving the speed of HPLC. We present a straightforward theoretical basis, informed by decades of literature on the problem of optimization, that is useful for comparing different technologies for improving the speed of HPLC. Through mindful optimization of conditions high performance separations on the sub-minute timescale are now possible and becoming increasingly common under both isocratic and gradient elution conditions, and the continued development of ultrafast separations will play an important role in the development of two-dimensional HPLC separations. Despite the relatively long history of HPLC as an analytical technique, there is no sign of a slow-down in the development of novel HPLC technologies.

Maximizing the peak capacity using coupled columns packed with 2.6μm core–shell particles operated at 1200bar

“…6-8. The shape of the KPL-curves in Fig. 5 corresponds very well to the theoretical expectations, according to which the kinetic performance limit curve (KPL-curve) should touch the Knox and Saleem limit (KS-limit) at the point corresponding to the optimal flow rate [3,10,31]. This can be understood as follows: adopting the LSStheory, the relation between the peak capacity and the number of theoretical plates generated by the column can be written as [22]:…”

“…The new generation of core-shell particles offers a quantum leap in chromatographic performance compared to their fully porous counterparts [1][2][3][4][5][6]. Although some of the reasons for this breakthrough are not fully understood yet [2], core-shell particles indisputably offer a reduced flow resistance ( 0 ) as well as an increased efficiency.…”

“…Combining these literature values and calculating the minimal separation impedance E min defined by Golay and Knox as [3,8,10]:…”

High Pressure Liquid Chromatography and Ultra‐high Pressure Liquid Chromatography of Plants: Basic Concepts

“…HPLC and UHPLC are still in full development, and column liquid chromatography is a method well prepared to cope with future analytical demands (Carr et al, 2011).…”

Streamlined approach to high‐quality purification and identification of compound series using high‐resolution MS and NMR

“…Nevertheless, an automated combination of results of both techniques supporting throughput applications has been missing. The strategy presented benefits from the recent developments in ultrafast HPLC applying sub-2 mm materials, high-resolution mass spectrometry (HRMS, TOF technology) confirming elemental composition with ppm accuracy as well as miniaturized NMR microprobes reducing compound consumption and time of analysis considerably [13,14].…”