Metabolites 2013, 3(3), 575-591; doi:10.3390/metabo3030575
Article

Combining Hydrophilic Interaction Chromatography (HILIC) and Isotope Tagging for Off-Line LC-NMR Applications in Metabolite Analysis

1email, 1email, 1,2email, 1email and 1,2,3,* email
Received: 17 May 2013; in revised form: 6 July 2013 / Accepted: 15 July 2013 / Published: 18 July 2013
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract: The complementary use of liquid chromatography (LC) and nuclear magnetic resonance (NMR) has shown high utility in a variety of fields. While the significant benefit of spectral simplification can be achieved for the analysis of complex samples, other limitations remain. For example, 1H LC-NMR suffers from pH dependent chemical shift variations, especially during urine analysis, owing to the high physiological variation of urine pH. Additionally, large solvent signals from the mobile phase in LC can obscure lower intensity signals and severely limit the number of metabolites detected. These limitations, along with sample dilution, hinder the ability to make reliable chemical shift assignments. Recently, stable isotopic labeling has been used to detect quantitatively specific classes of metabolites of interest in biofluids. Here we present a strategy that explores the combined use of two-dimensional hydrophilic interaction chromatography (HILIC) and isotope tagged NMR for the unambiguous identification of carboxyl containing metabolites present in human urine. The ability to separate structurally related compounds chromatographically, in off-line mode, followed by detection using 1H-15N 2D HSQC (two-dimensional heteronuclear single quantum coherence) spectroscopy, resulted in the assignment of low concentration carboxyl-containing metabolites from a library of isotope labeled compounds. The quantitative nature of this strategy is also demonstrated.
Keywords: metabolite profiling; metabolomics; NMR; HILIC; urine; 15N isotope tagging
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MDPI and ACS Style

Appiah-Amponsah, E.; Owusu-Sarfo, K.; Gowda, G.N.; Ye, T.; Raftery, D. Combining Hydrophilic Interaction Chromatography (HILIC) and Isotope Tagging for Off-Line LC-NMR Applications in Metabolite Analysis. Metabolites 2013, 3, 575-591.

AMA Style

Appiah-Amponsah E, Owusu-Sarfo K, Gowda GN, Ye T, Raftery D. Combining Hydrophilic Interaction Chromatography (HILIC) and Isotope Tagging for Off-Line LC-NMR Applications in Metabolite Analysis. Metabolites. 2013; 3(3):575-591.

Chicago/Turabian Style

Appiah-Amponsah, Emmanuel; Owusu-Sarfo, Kwadwo; Gowda, G.A. N.; Ye, Tao; Raftery, Daniel. 2013. "Combining Hydrophilic Interaction Chromatography (HILIC) and Isotope Tagging for Off-Line LC-NMR Applications in Metabolite Analysis." Metabolites 3, no. 3: 575-591.

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