A capillary electrophoresis coupled to mass spectrometry pipeline for long term comparable assessment of the urinary metabolome.
Although capillary electrophoresis coupled to mass spectrometry (CE-MS) has potential application in the field of metabolite profiling, very few studies actually used CE-MS to identify clinically useful body fluid metabolites. Here we present an optimized CE-MS setup and analysis pipeline to reproducibly explore the metabolite content of urine. We show that the use of a beveled tip capillary improves the sensitivity of detection over a flat tip. We also present a novel normalization procedure based on the use of endogenous stable urinary metabolites identified in the combined metabolome of 75 different urine samples from healthy and diseased individuals. This method allows a highly reproducible comparison of the same sample analyzed nearly 130 times over a range of 4 years. To demonstrate the use of this pipeline in clinical research we compared the urinary metabolome of 34 newborns with ureteropelvic junction (UPJ) obstruction and 15 healthy newborns. We identified 32 features with differential urinary abundance. Combination of the 32 compounds in a SVM classifier predicted with 76% sensitivity and 86% specificity UPJ obstruction in a separate validation cohort of 24 individuals. Thus, this study demonstrates the feasibility to use CE-MS as a tool for the identification of clinically relevant urinary metabolites.
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.
Franck
F
Boizard
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.
Valérie
V
Brunchault
HybridStat Predictive Analytics, Athens, Greece.
Panagiotis
P
Moulos
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.
Benjamin
B
Breuil
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.
Julie
J
Klein
Unité de Recherche Clinique Pédiatrique, Module Plurithématique Pédiatrique, Centre d'Investigation Clinique - Hôpital des Enfants, Toulouse, France.
Nadia
N
Lounis
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.
Cécile
C
Caubet
CHU Toulouse, Hôpital des Enfants, Service de Néphrologie - Médecine Interne - Hypertension Pédiatrique, Toulouse, France.
Stéphanie
S
Tellier
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.
Jean-Loup
JL
Bascands
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.|CHU Toulouse, Hôpital des Enfants, Service de Néphrologie - Médecine Interne - Hypertension Pédiatrique, Toulouse, France.
Stéphane
S
Decramer
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.
Joost P
JP
Schanstra
Institut National de la Santé et de la Recherche Médicale (INSERM), U1048, Institut of Cardiovascular and Metabolic Disease, Equipe 12, 1 avenue Jean Poulhès, BP 84225, 31432 Toulouse Cedex 4, France.|Université Toulouse III Paul-Sabatier Toulouse, France.
Bénédicte
B
Buffin-Meyer
D000328:Adult; D019075:Electrophoresis, Capillary / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D055442:Metabolome; D055432:Metabolomics / Q000379:methods*; D008875:Middle Aged; D014517:Ureteral Obstruction / Q000652:urine*
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