Broad-spectrum separations in metabolomics using enhanced polar LC stationary phases: A dedicated evaluation using plant metabolites

• Published in: Journal of separation science Vol. 30; no. 13; pp. 2002 - 2011
• Main Authors: t'Kindt, Ruben, Alaerts, Goedele, Vander Heyden, Yvan, Deforce, Dieter, Van Bocxlaer, Jan
• Format: Journal Article
• Language: English
• Published: Weinheim Wiley-VCH Verlag 01-08-2007; WILEY-VCH Verlag; WILEY‐VCH Verlag; Wiley
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical, structural and metabolic biochemistry; Arabidopsis - chemistry; Arabidopsis - metabolism; Biological and medical sciences; Chemical constitution; Chromatography, Liquid - instrumentation; Chromatography, Liquid - methods; Data Interpretation, Statistical; Extended polarity reversed-phase material; Fundamental and applied biological sciences. Psychology; Mass spectrometry; Mass Spectrometry - instrumentation; Mass Spectrometry - methods; Materials Testing; Microliquid chromatography; Plant metabolomics; Plant physiology and development; Proteome - analysis; Reproducibility of Results; Chemical analysis; Metabolite; Coupled method; HPLC chromatography; Extended polarity reversed-phase material; Arabidopsis thaliana; Reversed phase chromatography; Analysis method; Plant metabolomics; Cruciferae; Dicotyledones; Metabonomics; Angiospermae; Spermatophyta; Experimental plant; Mass spectrometry; Bonded stationary phase; Microliquid chromatography
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.200700077

In metabolomics, major efforts are invested in the development of suitable analytical approaches. A tendency towards the use of LC-MS is nowadays very obvious. A great majority of the metabolites of interest are polar to highly polar in nature. We focus on the so-called 'extended polarity' reversed LC phases, developed specifically to allow better retention characteristics for polar compounds. Several of these phases (Atlantis dC18, Inertsil ODS-3, Zorbax XDB, Alltima HP C18) are tested for different column dimension variations (0.5, 1.0, 2.1 mm id) in a specific LC-MS metabolomics setting. Important chromatographic and mass spectrometric quality parameters such as capacity factor, separation efficiency, peak symmetry, sensitivity, and mass accuracy are taken into account. All phases show adequate retention of polar compounds and also perform well with highly aqueous mobile phase compositions. On comparing 1.0 and 2.1 mm id columns, it is clear that the potential gain in sensitivity is not achieved. Using a Lockspray® device, accurate mass measurement with a Q-TOF micro is feasible within a mass range of 12 ppm if signal intensities of compound and lockmass are equated. Finally, the extended polarity RP approach in metabolomics experiments is endorsed using real plant extracts.