Substantial Benefits of an Inert Biphenyl Column for the Analysis of Steroids and Their Phase II Metabolites in Biological Samples
Author(s): Mathieu Galmiche1,2,3, Marie-Anaïs Monat1,2, Diego A. Lopez4, Cyrille Lamboley4, Paul Connolly4, Sergey Girel1,2, Davy Guillarme1,2, Isabel Meister1,2,3, Serge Rudaz1,2,3
1. School of Pharmaceutical Sciences, University of Geneva, Geneva, Switzerland, 2. Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, Geneva, Switzerland, 3. Swiss Centre for Applied Human Toxicology (SCAHT), Basel, Switzerland, 4. Restek Corporation,
Published By: Journal of Separation Science
Issue: 16, Volume 47
Year of Publication: 2024
Link: https://doi.org/10.1002/jssc.202400436
Abstract: Steroids can be used as biomarkers in clinical metabolomics and other fields related to human toxicology. This chemical group is known for its complexity, considering its number of isobaric compounds and the wide variety of phases I and II metabolic pathways that parent compounds can undergo. For a successful analysis of steroids in biological samples, liquid chromatography separation must be finely tuned. It is especially challenging for glucuronidated and sulfated steroids derivatives that bear polar heads and can be affected by non-specific adsorption. The benefits of a biphenyl stationary phase chemistry for the selectivity of the separation of steroids and their phase II metabolites and the extent to which nonspecific adsorption phenomena could degrade chromatographic performance were investigated. Replacing a conventional hardware by a passivated hardware allowed to considerably reduce peaks width and asymmetry of sulfated species. The addition of weak ion pairing agents in the mobile phase could also help to reduce non-specific adsorption but are detrimental to mass spectrometry detection. As confirmed by the successful detection of 52 steroids in plasma, the use of a biphenyl stationary phase complemented by a passivated column hardware is of great help for a successful biomedical analysis of steroids and their phase II metabolites.
Acknowledgment(s): This analysis features Restek’s Raptor Inert Biphenyl LC column (cat.# 9309A12-T). Learn more about our inert LC column technology.