*2.5. Statistical Analysis*

Experimental data were expressed as mean ± standard deviation (SD) of four biological replicates. Statistical analysis was performed by one-way analysis of variance (ANOVA), and Tukey's test was applied to establish significant differences among mean values at *p* < 0.05, using the Minitab 19.0 software package (State College, PA, USA). For multivariate analysis, raw data were normalized by the internal standard area, processed using log transformation (log 2), mean-centered, and divided by the square root of the deviation of each variable (Pareto scaling). Principal component analysis (PCA), heatmaps, and fold-change analysis were executed to evaluate differences between treated and untreated groups, using the MetaboAnalyst 4.0 server (https://www.metaboanalyst.ca accessed on 15 April 2021) [21].
