A hydrophilic and reversed-phase (HILIC-RP) liquid chromatography system for biomarkers in urine

What is it about?

Here it is described a two-dimensional liquid chromatographic system (2D-LC) based on hydrophilic interaction chromatography and reversed-phase (HILIC-RP) liquid chromatography with detection by mass spectrometry (MS) for the simultaneous separation of endogenous metabolites of clinical interest and excreted xenobiotics deriving from human exposure to toxic compounds The present proposal has the novelty of incorporating an on-line sample treatment based on the use of restricted access materials (RAMs), which permitted the direct injection of urine samples into the system. The work is focused on the instrumental coupling, studying all possible options and attempting to circumvent the problems of solvent incompatibility between the RAM device and the two chromatographic columns, HILIC and RP. The complete instrumental configuration developed, RAM-HILIC-RPLC-MS/MS, allowed the simultaneous assessment of urinary metabolites of clinical interest and excreted compounds derived from exposure to toxic agents with minimal sample manipulation. Thus, it may be of interest in areas such as occupational and environmental toxicology in order to explore the possible relationship between both types of compounds.

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Why is it important?

The main aim of this work was the development of a 2D-LC system based on HILIC and RPLC to simultaneously evaluate exogenous compounds (used as biomarkers of exposure to toxic agents) and endogenous metabolites of clinical usefulness as biomarkers of different pathologies, such as cancer and other diseases. The simultaneous evaluation of both types of compounds would be helpful to explore the possible relationship between exposure to toxic compounds and the levels of urinary metabolites related to the development of certain diseases. The establishment of this type of correlation could be of clinical interest for preventive purposes in areas such as occupational and environmental toxicology.