title: Integrated, nontargeted ultrahigh performance liquid chromatography/electrospray ionization tandem mass spectrometry platform for the identification and relative quantification of the small-molecule complement of biological systems.
abstract: To address the challenges associated with metabolomics analyses, such as identification of chemical structures and elimination of experimental artifacts, we developed a platform that integrated the chemical analysis, including identification and relative quantification, data reduction, and quality assurance components of the process. The analytical platform incorporated two separate ultrahigh performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS(2)) injections; one injection was optimized for basic species, and the other was optimized for acidic species. This approach permitted the detection of 339 small molecules, a total instrument analysis time of 24 min (two injections at 12 min each), while maintaining a median process variability of 9%. The resulting MS/MS(2) data were searched against an in-house generated authentic standard library that included retention time, molecular weight (m/z), preferred adducts, and in-source fragments as well as their associated MS/MS spectra for all molecules in the library. The library allowed the rapid and high-confidence identification of the experimentally detected molecules based on a multiparameter match without need for additional analyses. This integrated platform enabled the high-throughput collection and relative quantitative analysis of analytical data and identified a large number and broad spectrum of molecules with a high degree of confidence.
authors.affiliation: Metabolon, Incorporated, 800 Capitola Drive, Suite 1, Durham, North Carolina 27713, USA.
authors.forename: Anne M
authors.initials: AM
authors.lastname: Evans
authors.forename: Corey D
authors.initials: CD
authors.lastname: DeHaven
authors.forename: Tom
authors.initials: T
authors.lastname: Barrett
authors.forename: Matt
authors.initials: M
authors.lastname: Mitchell
authors.forename: Eric
authors.initials: E
authors.lastname: Milgram
mesh_terms: D000293:Adolescent; D000328:Adult; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D008297:Male; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D055815:Young Adult