title: HPLC-Based Metabolomic Analysis of Normal and Inflamed Gut.
abstract: The idiopathic inflammatory bowel diseases, which include Crohn's disease and ulcerative colitis, are multifactorial chronic conditions that result in numerous perturbations of metabolism in the gastrointestinal mucosa. Thus, methodologies for the qualitative and quantitative analysis of small molecule metabolites in mucosal tissues are important for further elucidation of mechanisms driving inflammation and the metabolic consequences of inflammation. High-performance liquid chromatography (HPLC) is a ubiquitous analytical technique that can be adapted for both targeted and non-targeted metabolomic analysis. Here, protocols for reversed-phase (RP) HPLC-based methods using two different detection modalities are presented. Ultraviolet detection is used for the analysis of adenine nucleotide metabolites, whereas electrochemical detection is used for the analysis of multiple amino acid metabolites. These methodologies provide platforms for further characterization of the metabolic changes that occur during gastrointestinal inflammation.
authors.affiliation: Department of Medicine, The University of Colorado School of Medicine, Aurora, CO, USA.|Mucosal Inflammation Program, The University of Colorado School of Medicine, Aurora, CO, USA.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Kao
authors.affiliation: Department of Medicine, The University of Colorado School of Medicine, Aurora, CO, USA.|Mucosal Inflammation Program, The University of Colorado School of Medicine, Aurora, CO, USA.
authors.forename: Jordi M
authors.initials: JM
authors.lastname: Lanis
authors.affiliation: Department of Medicine, The University of Colorado School of Medicine, Aurora, CO, USA.|Mucosal Inflammation Program, The University of Colorado School of Medicine, Aurora, CO, USA.
authors.forename: Erica
authors.initials: E
authors.lastname: Alexeev
authors.affiliation: Department of Microbiology and Immunology, Montana State University, Lewis Hall 109, Bozeman, MT, 59717, USA. douglas.kominsky@montana.edu.
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Kominsky
mesh_terms: D000225:Adenine / Q000032:analysis; D000596:Amino Acids / Q000032:analysis; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003106:Colon / Q000378:metabolism* / Q000473:pathology; D055664:Electrochemical Techniques; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000378:metabolism; D055432:Metabolomics / Q000379:methods*
chemical_list: D000596:Amino Acids; D000225:Adenine
keywords: Adenosine; Electrochemical detection; High-performance liquid chromatography; Inflammatory bowel disease; Intestinal epithelial cell; Tryptophan