Potential of HILIC‐MS in quantitative bioanalysis of drugs and drug metabolites

Abstract

One fundamental requirement for many lead optimization processes is the need for bioanalytical support within pharmaceutical drug discovery and development. Currently, most bioanalytical methods for pharmaceutical analysis employ HPLC coupled with MS/MS. The combination of HPLC and MS/MS detection frequently offers the complete resolution of the dosed compounds from their metabolites and the endogenous interferences to avoid extra efforts for chemical separation and sample clean-up procedures resulting in higher-throughput assays for a series of new chemical entities (NCEs). Hydrophilic interaction chromatography (HILIC) has been demonstrated to be a powerful technique for the retention of polar analytes offering a difference in selectivity compared to traditional RP chromatography. This review summarizes the HILIC-MS/MS methods for the trace quantitative determinations of the drug compounds and their metabolites to support both in vitro and in vivo experiments. The challenges on performing HILIC-MS/MS assays such as matrix ionization suppression and the potential for endogenous interferences are also presented.

Analytical Methods for Quantification of Drug Metabolites in Biological Samples

“…An appropriate amount of water (usually 5-15%) in the mobile phase is suggested for maintaining a stagnant enriched water layer on the surface of the polar stationary phase where the analytes partite. HILIC separates compounds by eluting with strong organic mobile phase against a hydrophilic stationary phase where elution is driven by increasing the water content in the mobile phase [83]. Although some column companies are marketing columns specific for HILIC, most columns used with normal phases, such as pure silica or cyano columns, can operate in HILIC conditions.…”

“…PGC chromatography commonly employs water, acetonitrile and methanol as the mobile phase but provides markedly greater retention and selectivity for polar analytes than reversed phase columns. For analyte elution PGC normally requires larger organic content in the mobile phase than reversed phase chromatography what consequently results in favorable sensitivity with MS detection [79,83,89].…”

Identifying and Overcoming Matrix Effects in Drug Discovery and Development

“…A relatively new column, the hydrophilic interaction LC (HILIC) column, consists of bare silica or a polar phase (amino, diol, cyano, etc.) bonded to silica and provides more favorable conditions for polar compound retention and ionization (Hsieh, 2008) (Ji et al, 2008). With HILIC chromatography, polar analytes have prolonged retention times and elute at high organic mobile phase content; therefore, they are separated from ion suppressing-early eluters and have high ionization efficiencies.…”

Theory and Practice of UHPLC and UHPLC–MS

“…An excellent overview on recent progress in the development of polar stationary phases until 2006 was written by Hemström and Irgum (2006); more recent advances are included in reviews by Jandera (2011) and by Buszewski and Noga (2012). Other reviews have focused on the effects of the operating conditions on HILIC separations (Hao et al, 2008), separation efficiency (Ikagami et al, 2008), HILIC method development (Dejaegher et al, 2008), coupling HILIC systems with MS and MS/MS (Nguyen and Schug, 2008;Hsieh, 2008), implementation of HILIC systems in two-dimensional (2D) separation modes (Jandera, 2008), and on HILIC applications in biological (Yoshida, 2004;Jian et al, 2010), pharmaceutical (Dejaegher and Vander Heyden, 2010), and metabolite (Iwasaki et al, 2007;Spagou et al, 2010) analysis. An excellent book on HILIC has appeared recently, covering various aspects of the technique: separation mechanism, stationary phases, method development, and applications (Olsen and Pack, 2013).…”