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  • 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

Sigma-Aldrich
D-(−)-Fructose, BioUltra, ≥99.0% (HPLC)
Sigma-Aldrich
L-Tyrosine, FG
Supelco
Xylitol, analytical standard
SAFC
Glycine
Sigma-Aldrich
Glycine, ACS reagent, ≥98.5%
Sigma-Aldrich
α-Ketoglutaric acid, 99.0-101.0% (T)
SAFC
L-Threonine
SAFC
L-Valine
SAFC
L-Tyrosine
Sigma-Aldrich
α-Ketoglutaric acid, BioReagent, suitable for cell culture, suitable for insect cell culture
Supelco
D-(−)-Fructose, analytical standard, analytical standard for fructose assay kit, for use with enzymatic assay kit FA20
Sigma-Aldrich
α-Ketoglutaric acid, ≥98.5% (NaOH, titration)
Supelco
Lactic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Malonic acid, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Mannitol, European Pharmacopoeia (EP) Reference Standard
Supelco
Mannitol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Methanol, suitable for HPLC, gradient grade, ≥99.9%
Sigma-Aldrich
L-Tyrosine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
L-Valine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Supelco
L-Valine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Supelco
L-Tyrosine, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-Threonine, reagent grade, ≥98% (HPLC)
USP
L-Tyrosine, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
Methanol, ACS reagent, ≥99.8%
Sigma-Aldrich
D-Mannitol, ≥98% (GC), suitable for plant cell culture
Sigma-Aldrich
D-Mannitol, BioXtra, ≥98% (HPLC)
Sigma-Aldrich
D-Mannitol, ≥98% (GC)
Supelco
D-Mannitol, ≥99.9999% (metals basis), for boron determination
Millipore
myo-Inositol, ≥99.0%, suitable for microbiology
Sigma-Aldrich
Glycine, 99%, FCC