Glycyrrhiza glabra L., also known as licorice, belongs to the Fabaceae family and is one of the most commercially valuable plants worldwide, being used in the pharmaceutical, cosmetic, and food industries, both for its therapeutic benefits as well as for the sweetening properties of the extract. This study evaluates the phytochemical composition, the biological activities, and the safety profile of a methanolic extract of licorice root (LRE) obtained from Romania. Ten phytocompounds were quantified by the HPLC-DAD-ESI+, the most abundant being the triterpene glycyrrhizin (13.927 mg/g dry extract.), followed by these flavonoids: liquiritin, liquiritigenin-apiosyl-glucoside, and apigenin-rutinoside liquiritigenin. The total phenolic content of the LRE was found to be 169.83 mg gallic acid/g dry extract. (GAE/g d.e.), and the extract showed a maximum of 79.29% antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Good antimicrobial activity of the LRE was observed for Gram-negative bacteria, especially for S. pneumoniae and S. pyogenes. The mineral content of the LRE was indicative of the lack of toxicity; heavy metals such as lead, cadmium, arsenic, nickel, and cobalt were below the detection limit. The safety profile of the licorice extract was assessed using the in vivo hen egg test-chorioallantoic membrane (HET-CAM protocol), indicating no irritability, good tolerability, and biocompatibility. The phytochemical and biological characterization of the Romanian licorice root extract reveals a good source of glycyrrhizin and polyphenols with antioxidant and antimicrobial potential, along with a safety profile that may be useful for future therapeutic applications.