LCGC North America-11-01-2015

During the course of my scientific career beginning in the 1960s, I have grown up with the birth of modern LC column technology, the refinements of the instrumentation, and the development of widespread application of this most powerful separation and analysis technique. In this installment, I would like to share with you some of my observations and experiences with the beginning, the growth period, and the maturation of HPLC columns, where I have focused nearly 33 years of writing for this magazine. I will explore some of the early column breakthroughs beginning with the development of large superficially porous particles (SPP), the porous irregular and spherical microparticulate particles, inorganic and organic polymeric monoliths and the rebirth of the current generation of SPP. In next month’s installment I will look into my crystal ball and see what the future of HPLC and UHPLC holds.

How to get started in the process of identifying the problem source for column-related problems.How to get started in the process of identifying the problem source for column-related problems.

So you think you know all about UV detection? Double check your understanding with this short primer on the principles and key variables of ultraviolet visible detection.

The capability to separate and analyze a wide range of proteins in complex systems remains a prime requirement in the biochemical sciences. Intact protein separations are especially difficult as these large molecules can present different conformations, association states and amphoteric features with chromatographic surfaces. Combining high performance liquid chromatography (HPLC) and ultrahigh pressure liquid chromatography (UHPLC) with mass spectrometry (MS) has proven to be an effective approach for solving difficult problems involving protein analyses. Considerable effort has been made to develop columns for separating proteins with high efficiency for reversed-phase, ion-exchange, size-exclusion chromatography, hydrophilic interaction liquid chromatography (HILIC), and hydrophobic interaction chromatography (HIC). Even so, many situations still exist where insufficient resolution is available for accurate protein analysis even when high-resolution MS is available. This presentation provides a brief overview of new approaches being investigated in the author's laboratories for obtaining superior protein separations. This includes new approaches for obtaining better protein separations with columns of highly-efficient superficially porous silica particles and techniques using MS-friendly mobile phases with effective methods for changing protein selectivity (band spacings) by column type and organic mobile phase modifiers.

The term “dead” volume often comes up in chromatography discussions and literature. This month, GC Connections addresses the nature of this phenomenon, when it can become a problem that affects chromatographic results, and how to understand and take control of it.

Click the title above to open the LCGC North America November 2015 regular issue, Vol 33 No 11, in an interactive PDF format.