In this paper, natural deep eutectic solvents (NADESs) with lactic acid, glycine, ammonium acetate, sodium acetate, and choline chloride were prepared with and without the addition of water. NADES formation was evaluated using FTIR and Raman, where hydrogen bonds were identified between the hydroxyl group of lactic acid and the amino and carboxyl groups of glycine. Acetate and ammonium ions were also identified as forming bonds with lactic acid. The addition of water did not cause changes in the vibrational modes of the FTIR and Raman spectra but contributed to a reduction in NADES viscosity and density. Viscosity ranged from 0.335 to 0.017 Pa s−1, and density ranged from 1.159 to 0.785 g mL−1. The best results for the extraction of phenolic compounds from pitaya (dragon fruit) were achieved with an organic solvent (450. 41 mg 100 g−1 dry bases-db) in comparison to NADESs lactic acid:glycine (193.18 mg 100 g−1 db) and lactic acid:ammonium acetate (186.08 mg 100 g−1 db). The antioxidant activity of the extracts obtained with the NADESs was not statistically different from that of the extract obtained with organic solvents.