List of triacylglycerols in liver identified with significant concentration changes between the control cohort and the test cohort at two and four weeks.

a<p>Fold-change is the ratio of average peak area of a metabolite in the test cohort (T) to that in the control cohort (C).</p>b<p>na refers to a metabolite that was detected only in the test cohort. Therefore, the values of fold change for these metabolites are not available.</p

Time course changes of hepatic lipid content.

<p>(A) Hepatic neutral lipid detected by Oil red O staining of cryostat liver sections. Alcohol exposure increased hepatic neutral lipid (lipid droplets) gradually along the 4 weeks of experiment. (B) Hepatic TG concentrations were quantitatively measured. Data are expressed as mean ± SD (<i>n</i> = 6−8). Statistical differences were analyzed by ANOVA followed by Bonferroni <i>post hoc</i> comparison, and means without a common letter differ at <i>p</i><0.05.</p

Body weight, liver weight and plasma parameters of the control cohort and the test cohort at two and four weeks.

<p>Data are expressed as mean ± SD (<i>n</i> = 6−8). Statistical differences were analyzed by ANOVA followed by Bonferroni <i>post hoc</i> comparison, and means without a common letter differ at <i>p</i><0.05.</p

Time course changes of WAT tissues.

<p>(A) WAT mass. The weights of both eWAT and sWAT in control mice increased gradually during the 4 weeks of experiment. However, the alcohol-fed mice did not show weight change in both eWAT and sWAT at either 2 weeks or 4 weeks. (B) WAT to body weight ratio (%). Data are expressed as mean ± SD (<i>n</i> = 6−8). Statistical differences were analyzed by ANOVA followed by Bonferroni <i>post hoc</i> comparison, and means without a common letter differ at <i>p</i><0.05.</p

Time course trajectory of triacylglycerol TG(16∶0/18∶2/20∶4)[iso6] without any deuterium labeling in liver samples.

<p>Time course trajectory of triacylglycerol TG(16∶0/18∶2/20∶4)[iso6] without any deuterium labeling in liver samples.</p

An example of identifying a deuterium incorporated metabolite using MS/MS information.

<p>The metabolite ion <i>m/z</i> value was measured on FTICR-MS as 904.74066. (A) is the experimental MS/MS spectrum of non-deuterated metabolite. (B) is the matching result of the non-deuterated metabolite between the experiment MS/MS spectrum and the theoretical MS/MS spectrum generated by <i>Mass Frontier</i>. The matched fragment ions are highlighted in red and the not matched ions in black. (C) is the MS/MS spectrum of deuterium incorporated metabolite.</p

Sample time course trajectories of deuterium labeled triacylglycerols detected in liver, eWAT and sWAT samples.

<p>(A) TG(16∶0/18∶2/20∶4)[iso6] with one ²H label and one Na⁺ as adduct ion in liver samples. (B) TG(16∶0/16∶1/16∶1)[iso3] with one ²H label and an adduct ion of Na⁺ eWAT samples. (C) TG(16∶0/16∶0/18∶1)[iso3] with one ²H label and an adduct ion of Na⁺ in sWAT samples.</p

Sample concentration changes of metabolite in two different physiological conditions.

<p>The abundance test (pair-wise two-tail t-test) shows that the concentration of this metabolite in the test cohort is increased with a fold change of 2.8 and a p-value of 1.4×10⁻⁵. This metabolite was further identified as TG(16∶0/18∶2/20∶4)[iso6] by MS/MS analysis.</p