title: Ultra high resolution SFC-MS as a high throughput platform for metabolic phenotyping: application to metabolic profiling of rat and dog bile.
abstract: Ultra high resolution SFC-MS (on sub-2μm particles) coupled to mass spectrometry has been evaluated for the metabolic profiling of rat and dog bile. The selectivity of the SFC separation differed from that seen in previous reversed-phase UPLC-MS studies on bile, with the order of elution for analytes such as e.g., the bile acids showing many differences. The chromatography system showed excellent stability, reproducibility and robustness with relative standard deviation of less than 1% for retention time obtained over the course of the analysis. SFC showed excellent chromatographic performance with chromatographic peak widths in the order of 3s at the base of the peak. The use of supercritical fluid carbon dioxide as a mobile phase solvent also reduced the overall consumption of organic solvent by a factor of 3 and also reduced the overall analysis time by a factor of 30% compared to reversed-phase gradient LC. SFC-MS appear complementary to RPLC for the metabolic profiling of complex samples such as bile.
authors.affiliation: Waters Corporation, Milford, MA, USA; Analytical & Environmental Sciences Division, King's College London, Stamford Street, London, UK.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Jones
authors.affiliation: Waters Corporation, Milford, MA, USA.
authors.forename: Paul D
authors.initials: PD
authors.lastname: Rainville
authors.affiliation: Waters Corporation, Milford, MA, USA.
authors.forename: Giorgis
authors.initials: G
authors.lastname: Isaac
authors.affiliation: Computational and Systems Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, UK.
authors.forename: Ian D
authors.initials: ID
authors.lastname: Wilson
authors.affiliation: Analytical & Environmental Sciences Division, King's College London, Stamford Street, London, UK.
authors.forename: Norman W
authors.initials: NW
authors.lastname: Smith
authors.affiliation: Computational and Systems Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, UK. Electronic address: r.plumb@imperial.ac.uk.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Plumb
mesh_terms: D000818:Animals; D001646:Bile / Q000737:chemistry* / Q000378:metabolism; D001647:Bile Acids and Salts / Q000032:analysis; D025924:Chromatography, Supercritical Fluid / Q000379:methods*; D004285:Dogs; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D055432:Metabolomics / Q000379:methods*; D025341:Principal Component Analysis; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results
chemical_list: D001647:Bile Acids and Salts
keywords: Bile acids; Bile metabolites; Metabolic profiling; Metabolomics; Metabonomics; SFC