*4.6. Statistical Analysis*

The data were analyzed using two-way ANOVA, with individual comparisons being made using Tukey's multiple comparison test. All analyses were performed using R [52]. Statistical significance was set at *p* < 0.05, and the data are expressed as the means ± standard error of the mean (SEM).

Quantitative enrichment analysis using the pathway-associated metabolite sets included in MetaboAnalyst 4.0 [53], which is an established tool for metabolite set enrichment analysis, was performed using the plasma metabolite compounds that were significantly a ffected by either the HT treatment or orotic acid supplementation. Di fferences were considered significant at *p* < 0.05.
