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Sea fennel is a halophytic plant rich in valuable nutritional components and is characterized by pleasant organoleptic properties. While its essential oils (EOs) are well investigated, there are no reports on the volatiles from their corresponding hydrolates, which are the main by-products of EO isolation, as well as on their biological activity. Therefore, the composition and biological activities of EOs and corresponding hydrolates of sea fennel from Atlantic (French, FRA) and Mediterranean (Croatian, CRO) ecotypes were investigated and compared. The EO from the CRO sample was characterized by an abundance of sabinene and limonene, while that from the FRA ecotype was rich in dillapiol and carvacryl methyl ether. The CRO hydrolate was rich in terpinen-4-ol and 10-(acetylmethyl)-3-carene, while dillapiol, thymyl methyl ether and γ-terpinene were the main compounds in the FRA sea fennel hydrolate. The biological activities of the EOs and hydrolates were evaluated for their antioxidant (with DPPH, NO, FRAP and ORAC bioassays), antimicrobial (against some Gram+ and Gram- spoilage bacteria) and antiageing (tyrosinase, elastase and collagenase inhibition) activities. Both EOs showed low reducing powers and antiradical activities while the ability of both hydrolates to quench NO was slightly higher (35–39% if inhibition). The FRA EO showed low activity against Staphylococcus aureus (8 mm), while CRO moderately inhibited the growth of P. aeruginosa (8 mm), but strongly inhibited the other two bacterial strains. While the French EO showed no antityrosinase and anticollagenase activity, the Croatian oil significantly inhibited both enzymes (IC50 of 650 µg/mL and IC50 of 2570 µg/mL, respectively) probably due to the dominance of limonene and sabinene. Neither EO exhibited antielastase properties, while the hydrolates from both ecotypes showed no antiageing activity, regardless of the enzyme tested. The EOs from the aerial parts of sea fennel from FRA and CRO differed greatly in composition, resulting in different activities. The Croatian samples appeared to have better biological properties and are therefore good candidates for applications as preservatives or antiageing agents. [ABSTRACT FROM AUTHOR]