LCGC Europe-03-01-2019

This article is an update on the technique of HILIC and covers recent ideas on the mechanism of separation, and how it may be manipulated to suit the separation of particular sample types. The advantages of HILIC are discussed, and also the actual and perceived disadvantages of the technique and how the latter can be overcome. Some new applications of HILIC for characterization of biopharmaceuticals, where it can even be applied to the separation of intact proteins, and to applications in metabolomics, will be discussed.

The offerings of commercially available columns for reversed-phase liquid chromatography (LC) continue to expand. Are these columns similar or different compared to what is already available?

Solid-phase microextraction (SPME) was introduced nearly 30 years ago and since that time has matured into a widely used tool in the arsenal of sample preparation techniques. Simultaneously, it has spawned a host of related techniques where sorbent coatings are placed on stir bars, magnetic particles, vial walls, and so on. Over the past few years, several advances in SPME have been developed, including increasing the sorbent surface area available for extraction, accommodating direct analysis by mass spectrometry (MS), and sorbent overcoating to resist fouling by sugars, lipids, and other macromolecules present in some sample types. These advances are discussed in this month’s instalment. The use of SPME for microsampling of biological systems, so-called bio-SPME, will be the focus of Part 2.

Click the title above to open the LCGC Europe March 2019 regular issue, Vol 32, No 03, in an interactive PDF format.

CZE–ESI‑TOF‑MS for the characterization of the mAb infliximab and its variants is presented. Infliximab was analyzed using a middle-up approach involving either reduction or digestion with the enzyme IdeS. A multilayer capillary coating of PB-DS‑PB in combination with a background electrolyte of 40% acetic acid provided efficient separation of the obtained antibody fragments, that is, LC and HC, as well as F(ab’)2 and Fc/2 parts. C-terminal lysine variants were also resolved. Recorded mass spectra of HC and Fc/2 fragments permitted assignment of 13 glycoforms and provided a quantitative profile, with G0F the most abundant glycoform (~50%). CZE–ESI-TOF-MS represents an efficient means for the straightforward analysis of a monoclonal antibody and its proteoforms.

One millimetre internal diameter liquid chromatography columns are available from many manufacturers. In this article, the utility of 1.0-mm internal diameter (i.d.) columns, and the arenas in which they play a relatively strong role, are investigated. Further, the advantages and disadvantages of 1.0 mm diameter columns are contrasted with both larger- and smaller-bore formats.

Ten years since its official definition, foodomics continues to expand the scientific knowledge of food and nutrition while resolving many analytical challenges along the way. LCGC Europe spoke to Alejandro Cifuentes from the Institute of Food Science Research, in Madrid, Spain, about his current foodomics research projects, the overall state of the field, and the future of foodomics.

The 48th International Symposium on High Performance Liquid Phase Separations and Related Techniques (HPLC 2019) will be held 16–20 June 2019 at the Milano-Bicocca University, Milan, Italy. This is the first time that this symposium will be held in Italy.

A discussion of hydrophobic interaction chromatography (HIC) theory and its application to the analysis of proteins and biomolecules is presented.

This information is supplementary to the article “Middle-Up Characterization of the Monoclonal Antibody Infliximab by Capillary Zone Electrophoresis-Mass Spectrometry” that was published in the March 2019 issue of LCGC Europe.