The genus Salvia has recently attracted great attention due to its notable biological activities. Within this context, in this study, the chemical characterization and biological effects of three extracts (dichloromethane (DCM), methanol (MeOH), and water) from three Salvia species (S. blepharochlaena (SB), S. euphratica var. leiocalycina (SE), and S. verticillata subsp. amasica (SV)) were assessed. For the chemical characterization, the qualitative and quantitative analysis of phenolic components in the methanol extracts was carried out by high-performance liquid chromatography with electrospray ionization mass spectrometry detection (HPLC-UV-ESI-MSn). Total phenolic, flavonoid and phenolic acid contents were also studied. Concerning the biological effects, antioxidant (DPPH and ABTS free radical scavenging; ferric (FRAP) and cupric (CUPRAC) reducing power; phosphomolybdenum, and metal chelating assays), enzyme inhibitory (cholinesterase, tyrosinase, amylase, glucosidase, lipase, and elastase) and cytotoxic effects (A-549 and MCF-7 cell lines) were evaluated. After the evaluation of the phytochemical profile by HPLC-UV-ESI-MSn, it was observed that the main compound in the analyzed extracts was rosmarinic acid, which was present at high concentrations, particularly in SV, which presented rosmarinic acid levels higher than the usual levels found in other Salvia species or related plants. Generally, the SV-water extract presented the strongest antioxidant abilities with higher levels of total bioactive compounds. However, the studied DCM extracts had higher enzyme inhibitory potentials compared with MeOH and water extracts. SE-DCM exerted the most potent cytotoxic effects, followed by SB-water and SB-MeOH extracts.