Column Chemistry

Martin Giera and colleagues from the Center of Proteomics andMetabolomics (The Netherlands) have published a study in the Journal of Chromatography B¹ evaluating different column chemistries for reversed-phase liquid chromatography (RP–LC) and hydrophilic interaction chromatography (HILIC) for fast urinary metabolic profiling.

Urine is often chosen as a sample because its collection offers the least discomfort to patients compared to blood or cerebrospinal fluid. However,it also contains greater levels of highly hydrophilic metabolites that can bechallenging analytically.

According to Giera, RP–LC material has dominated the field up untilrecently, but HILIC is becoming increasingly recognized as an alternative orcomplementary column chemistry. Giera told The Column: “If a column cannot sufficiently retain certain metabolites, various negative effects like ionization suppression will play a large role.”

The levels of 54 common urinary metabolites in pooled urine sampleswere analysed. The materials selected for testing were core–shell C18-,XB-C18-, and PFP-RPLC; and three porous HILIC columns: amino, cyano,and diol. Giera commented that analysing and evaluating the resultant30,000 generated signals was challenging. It was addressed by applying“designated computer scripts” and a “generic peak scoring algorithm.”

Giera told The Column: “Our study strongly suggests the use of HILICand particularly diol-HILIC columns for urinary metabolic profiling as moremolecular information can be obtained in comparison to RPLC columns.”

Reference

1. M. Giera et al, Journal of Chromatography B927, 90–96 (2013).

This story originally appeared in The Column. Click here to view that issue.