The detection of phenotypic differences in the metabolic plasma profile of three strains of Zucker rats at 20 weeks of age using ultra‐performance liquid chromatography/orthogonal acceleration time‐of‐flight mass spectrometry

Abstract

Analysis of biological fluids using ultra-performance liquid chromatography/mass spectrometry (UPLC/MS) (metabonomics) can allow new insights to be gained into disease processes, with advances in chromatographic techniques enabling the detection of thousands of metabolites. In this work metabonomics has been used to investigate the metabolic processes involved in type II diabetes in the Zucker obese rat. Plasma was analyzed from three different strains, the Zucker (fa/fa) obese, Zucker lean and the lean/(fa) obese cross. Using UPLC/MS, ca. 10,000 ions were detected due to the narrow peak widths and excellent peak shapes achieved with this technology. Confidence in the chromatographic performance was demonstrated by the use of quality control standards. The positive and negative ion total ion chromatograms obtained from the three strains were readily distinguishable using multivariate statistical analysis. The greatest difference was observed between the Zucker lean and Zucker lean/(fa) rats compared to the Zucker (fa/fa) obese rats. Positive ions m/z 220 (4.36 min), 282(3.78 min), 359 (5.33 min) and 405 (7.77 min) were elevated in the plasma derived from Zucker lean rats whilst ions m/z 385 (6.80 min) and 646 (4.36 min) were at a lower concentration compared to the plasma from the Zucker (fa/fa) obese animals. Negative ions elevated in the Zucker lean rats included m/z 212 (2.30 min), 514 (2.85 min), 295 (4.39 min), 329 (3.11 min), 343 (2.86 min) and 512 (2.86 min) with ions m/z 538 (4.18 min), 568 (4.18 min), 568 (5.09 min) and 612 (4.30 min) being raised in the samples derived from Zucker (fa/fa) obese animals. The ion m/z 514 (3.85 min) was found to correspond to taurocholate, providing further support for an involvement of taurine metabolism in diabetes.

Fundamental and practical aspects of ultrahigh pressure liquid chromatography for fast separations

“…The peak capacity and the number of peaks detected using fast UPLC gradients and TOF-MS were similar to, or better than, that generated with a 10 min LC separation. The same group used UPLC/MS to detect phenotypic differences in the metabolic plasma profile of three strains of Zucker rats [91]. They were able to detect approximately 10 000 ions from the chromatogram due to the narrow peak widths and excellent peak shapes achieved with this technology.…”

Metabonomics in Diabetes Research

“…53 Again taurocholate was found to be raised in obese Zucker (fa/fa) rats. The additional markers detected in this study remained unidentified; however, they allowed a metabolic class separation of the three rat strains investigated.…”

Metabolomic profiling of biomarkers of liver X receptor-induced toxicity in mouse liver tissue

“…Mass spectrometry has also been employed extensively for metabolomics studies (Plumb et al 2002(Plumb et al , 2003(Plumb et al , 2006(Plumb et al , 2008(Plumb et al , 2009Idborg-Bjorkman et al 2003;Lafaye et al 2003;Lenz et al 2004a, b;Wilson et al 2005;Dettmer et al 2007) as it offers improved sensitivity over NMR which is useful for untargeted studies to identify novel biomarkers. Mass spectrometry is frequently coupled with a prior separation technique such as HPLC, UPLC, Capillary LC or GC, e.g.…”