Skip to Content
Merck
  • Urinary metabolic fingerprinting of mice with diet-induced metabolic derangements by parallel dual secondary column-dual detection two-dimensional comprehensive gas chromatography.

Urinary metabolic fingerprinting of mice with diet-induced metabolic derangements by parallel dual secondary column-dual detection two-dimensional comprehensive gas chromatography.

Journal of chromatography. A (2014-08-28)
Davide Bressanello, Erica Liberto, Massimo Collino, Stephen E Reichenbach, Elisa Benetti, Fausto Chiazza, Carlo Bicchi, Chiara Cordero
ABSTRACT

This study investigates the potential of a parallel dual secondary column-dual detection two-dimensional comprehensive GC platform (GC×2GC-MS/FID) for metabolic profiling and fingerprinting of mouse urine. Samples were obtained from a murine model that mimics a typical unhealthy Western diet featuring both high fat and sugar (HFHS) intake, which induces obesity, dyslipidemia, and insulin resistance. Urines collected at different steps of the study were used to obtain pivotal and comparative data on the presence and relative distributions of early markers of metabolic disease. The data elaboration and interpretation work-flow includes an advanced untargeted fingerprinting approach, with peak-region features to locate relevant features to be quantified by external standard calibration. The reliability of untargeted fingerprinting is confirmed by quantitative results on selected relevant features that showed percentages of variations consistent with those observed by comparing raw data quantitative descriptors (2D peak-region volumes and percent of response). Analytes that were up-regulated with % of variation ranging from 30 to 1000, included pyruvic acid, glycerol, fructose, galactose, glucose, lactic acid, mannitol and valine. Down-regulation was evidenced for malonic acid, succinic acid, alanine, glycine, and creatinine. Advanced fingerprinting also is demonstrated for effectively evaluating individual variations during experiments, thus representing a promising tool for personalized intervention studies. In this context, it is interesting to observe that informative features that were not discriminant for the entire population may be relevant for individuals.

MATERIALS
Product Number
Brand
Product Description

Supelco
Acetic anhydride, derivatization grade (GC derivatization), LiChropur, ≥99.0%
USP
Malic acid, United States Pharmacopeia (USP) Reference Standard
Supelco
Creatinine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Glycerol, ReagentPlus®, ≥99.0% (GC)
Sigma-Aldrich
Dichloromethane, ACS reagent, ≥99.5%, contains 40-150 ppm amylene as stabilizer
Sigma-Aldrich
Dichloromethane, ACS reagent, ≥99.5%, contains 40-150 ppm amylene as stabilizer
Supelco
Succinic Acid, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Malic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Dichloromethane, contains 40-150 ppm amylene as stabilizer, ACS reagent, ≥99.5%
Sigma-Aldrich
Acetic anhydride, Arxada quality, ≥99.5% (GC)
Supelco
Gallic acid, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
DL-Malic acid, meets analytical specification of FCC, E296, 99-100.5% (alkalimetric)
Supelco
N-Methyl-N-(trimethylsilyl)trifluoroacetamide, BioReagent, for silylations, LiChropur
Supelco
N-Methyl-N-(trimethylsilyl)trifluoroacetamide, synthesis grade
Sigma-Aldrich
L-Alanine-12C3, 99.9 atom % 12C
Supelco
N-Methyl-N-(trimethylsilyl)trifluoroacetamide, derivatization grade (GC derivatization), LiChropur, ≥98.5%
Sigma-Aldrich
Dichloromethane, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%, contains 50-150 ppm amylene as stabilizer
Sigma-Aldrich
Hexane, Laboratory Reagent, ≥95%
Sigma-Aldrich
Hexane, ReagentPlus®, ≥99%
Sigma-Aldrich
Hexane, suitable for HPLC, ≥95%
Sigma-Aldrich
Acetic anhydride, ACS reagent, ≥98.0%
Sigma-Aldrich
Acetic anhydride, ReagentPlus®, ≥99%
Sigma-Aldrich
Acetic anhydride, 99.5%
Supelco
Pyridine, analytical standard
Sigma-Aldrich
Pyridine, suitable for HPLC, ≥99.9%
Supelco
Pyridine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Pyridine, ≥99%
Sigma-Aldrich
Pyridine, biotech. grade, ≥99.9%
Sigma-Aldrich
Pyridine, ACS reagent, ≥99.0%
Sigma-Aldrich
Succinic acid, BioReagent, BioRenewable, suitable for cell culture, suitable for insect cell culture