Background Halophytes are considered as valuable sources of traditional drugs in different countries. Purpose The present study aimed to evaluate biological and chemical fingerprints of three halophytes (Arthrocnemum macrostachyum (Moric.) C, Koch, Halimione portulacoides (L.) Aellen and Salicornia europaea L.). Materials and methods The antioxidant and enzymatic inhibitory potential (acetylcholinesterase, butyrylcholinesterase, α-amylase, α-glucosidase, and tyrosinase) were assessed. The total phenolic, flavonoid contents, and the chemical profiles were appraised using the ultra-high performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Molecular docking was conducted to provide additional insights of molecular interactions of the enzymes/phytochemicals. Results Ethyl acetate extract was the most efficient extract, with A. macrostachyum being the most potent towards DPPH and ABTS radicals and phosphomolybdenum assay. Ethyl acetate extract of A. macrostachyum was also the best reducing agent (CUPRAC and FRAP assays). Methanol and ethyl acetate extract of A. macrostachyum, H. portulacoides, and S. europaea showed significant enzyme inhibition potential. Ethyl acetate extract of A. macrostachyum showed the highest total phenolic (29.54 ± 0.78 mgGAEs/g extract) while the ethyl acetate extract of S. europaea was more abundant in flavonoids (18.26 ± 0.11 mgREs/g extract). Phytochemical profiling allowed the identification of several components in the methanolic extracts (16 in A. macrostachyum, 14 in H. portulacoides, and 11 in S. europaea), with quinic acid, p-coumaric acid, and rhamnetin being most abundant. Docking studies revealed that the above compounds showed scores for the enzymes tested. Conclusion The three halophytes studies could be considered as potential sources of biologically-active compounds for novel phytopharmaceuticals development.