Hydrophilic interaction chromatography (HILIC) is a relatively recently introduced mode of liquid-phase separations. Recently, HILIC has been used for coupling to MS in metabonomic/metabolomic studies to provide a complementary tool to the widely used reversed-phase (RP) chromatographic separations. The combination of HILIC with MS detection covers a number of polar metabolites that are typically nonretained in RPLC-mode separations and thus enlarging the number of detected analytes. This way of metabolite profiling thus provides more comprehensive metabolome coverage than using RP chromatography alone. This review describes the applications and the utility of HILIC-MS in metabolomic/metabonomic studies and highlights certain characteristic examples in the life and plant-food sciences.
Hydrophilic interaction ultra performance liquid chromatography (HILIC-UPLC) permits the analysis of highly polar metabolites, providing complementary information to reversed-phase (RP) chromatography. HILIC-UPLC-TOF-MS was investigated for the global metabolic profiling of rat urine samples generated in an experimental hepatotoxicity study of galactosamine (galN) and the concomitant investigation of the protective effect of glycine. Within-run repeatability and stability over a large sample batch (>200 samples, 60 h run-time) was assessed through the repeat analysis of a quality control sample. Following system equilibration, excellent repeatability was observed in terms of retention time (CV < 1.7%), signal intensity (CV < 14%), and mass variability (<0.005 amu), providing a good measure of reproducibility. Classification of urinary metabolic profiles according to treatment was observed, with significant changes in specific metabolites after galN exposure, including increased urocanic acid, N-acetylglucosamine, and decreased 2-oxoglutarate. A novel finding from this HILIC-UPLC-MS approach was elevated urinary tyramine in galN-treated rats, reflecting disturbed amino acid metabolism. These results show HILIC-UPLC-MS to be a promising method for global metabolic profiling, demonstrating high within-run repeatability, even over an extended run time. Retention of polar endogenous analytes and xenobiotic metabolites was improved compared with RP studies, including galN, N-acetylglucosamine, oxoglutarate, and urocanic acid, enhancing metabolome coverage and potentially improving biomarker discovery.
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